Objective:To study the mutation frequencies of the exon 5 and the exon 8 of PTEN (phosphatase and tensin homology deleted on chromosome ten) gene in gastric carcinoma and investigate the relationship of the gene mutation and pathological differentiation and clinical stage.Methods:The mutation of exon 5 and exon 8 of PTEN gene was detected in 42 gastric carcinoma samples and the matched adjacent normal gastric mucosa with polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP) method.The PCR products of mutant samples were analysed by DNA sequencing technique.Results:The mutation of PTEN was shown in 3 of the 42 gastric carcinoma tissues and in none of the adjacent normal tissues.The mutation rates of PTEN gene in poorly differentiated and well differentiated samples were 12.00% and 0,respectively (P0.05).The mutation rates of PTEN gene in clinical stage Ⅰand Ⅱ (5.88%) had no significant difference with that in clinical stage Ⅲand Ⅳ (8.00%) (P0.05).Conclusion:PTEN gene mutation occurs mainly in poorly differentiated gastric carcinoma tissues,and the mutation rate is not related to pathological differentiation and clinical stage.

As a depot drug delivery system, injectable polylactide-polyglycolide (PLGA) sustained-release microspheres have been successfully used to treat many diseases since the first microsphere product Lupron depot was approved for marketing in the United States in 1989. It has the ability of long-term release in the body for several days to several months, which can not only reduce the times of administration, but also reduce the drug blood concentration fluctuations, significantly improve the safety and patient compliance. In vitro-in vivo correlation (IVIVC) makes the development of microspheres more possible. It can describe the dynamic information of drug release in vivo through the in vitro release behavior of microspheres, and can reduce the workload of each stage and shorten the time span while characterizing the performance of microspheres. IVIVC can provide guidance or support for drug development, production changes, supervision and management. This article summarizes the release mechanism of injectable PLGA sustained-release microspheres, common measurement methods and theories of in vitro and in vivo release. And we also focus on the establishment and application of IVIVC, especially A level IVIVC in the field of microsphere preparations, to provide a reference for further study on in vitro-in vivo correlation of microspheres.

Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P

AIM:To evaluate the application and effect of bandage contact lens in pterygium excision combined with conjunctival transplantation (CAT).METHODS:In a prospective, randomized, controlled clinical study, 110 patients (110 eyes) diagnosed with primary pterygium were collected in PLA NO.474 Hospital from January 2015 to January 2016.The 110 patients enrolled in the study in turn, and divided into two groups by the odd and even number.The odd number divided into bandage contact lens group (CAT + bandage contact lens, n=55), while the even number divided into control group (CAT, n=55).Visual analog scale (VAS) and corneal irritation were evaluated on 1, 3 and 7d after operation.Cornea fluorescent staining testing was carried out on 3d after operation.Following-up all the patients with 1a at least observed the recurrence of pterygium.RESULTS:The score of VAS in bandage contact lens group less than that in control group on 1d (4.13±2.06 vs 5.80±1.93, t=4.391, P<0.001) and 3d (2.09±1.36 vs 3.65±1.65, t=5.422, P<0.001) after operation, while there was no significant difference between two groups on 7d (t=1.295, P=0.198) after operation.The corneal irritation in bandage contact lens group less than that in control group on 1d and 3d after operation (P<0.05), while there was no significant difference between two groups on 7d after operation (P=0.052).Cornea fluorescent staining testing area in bandage contact lens group was less than that in control group on 7d after operation (0.33±0.37mm2 vs 2.73±2.21mm2, t=7.921, P<0.01).There was no significant difference in recurrence rate between two groups after 1a operation (P=1.000).CONCLUSION:Bandage contact lens could significantly release pain and corneal irritation, promote the healing of the corneal epitheliums in the defected area, and increase the postoperative comfort level in patients after operation.

Objective:To establish a comprehensive and effective scoring model based on ultrasonic characteristics for predicting the restenosis risk after superficial femoral artery stenting, in order to assess the possibility of in-stent restenosis and to provide guidance for the selection of therapeutic strategies.Methods:A retrospective review of a database of 328 patients (381 limbs) undergoing superficial femoral artery stents in Xuanwu Hospital, Capital Medical University from January 2016 to January 2018 was made as a modeling group.In the modeling cohort, the multivariate logistic regression analysis was performed to screen independent risk factors for in-stent restenosis. A predictive scoring model of restenosis risk was established with weighted score of independent risk factors according to the odd ratio values. Based on the best cut-off value of the receiver operating characteristic (ROC) curves, the scoring table was divided into low-risk and high-risk groups of restenosis.Results:Multivariate logistic regression analysis showed that 8 factors were included in the score system to establish the scoring model of in-stent restenosis risk prediction including calcified plaque, peak systolic velocity of popliteal artery<40 cm/s, runoff scores≥4, ankle-brachial index<0.5, female (1 point each); complicated stroke, complicated chronic renal disease, total lesion length 15.0-24.9 cm (2 points each); total lesion length≥25.0 cm (3 points), a total of 12 points in the model. The validation indicated that the scoring system had good predictive value(AUC=0.775, 95% CI=0.727-0.824, P<0.001) and goodness of fit (Hosmer-Lemeshow χ 2=4.921, P=0.766). The agreement with digital subtraction angiography(DSA) was good (Kappa value=0.609). The scoring system was further divided into the low-risk restenosis (0-5 points) and high-risk restenosis (6-12 points) according to the best cut-off value of 5.5, with a sensitivity of 68.1%, a specificity of 74.6%, and the accuracy of 72.7%. Conclusions:The superficial femoral artery in-stent restenosis risk predicting score model based on ultrasonic characteristics may accurately predict the restenosis preoperatively. It provides a theoretical basis for the precise surgical plans.

Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.

OBJECTIVE To investigate how MLKL functions on the membrane and explore its electrophysiological characters and structure. METHODS The full-length human MLKL were expressed in SF21 cells and purified using glutathione-sepharose affinity chromatography. The currents of purified MLKL proteins were recorded in avoltage-clamp mode using a Warner BC-535 bilayer clamp amplifier. The currents were digitized using pCLAMP 10.2 software. HEK293 cells were cultured and transfected with MLKL plasmid. Cell viability was examined using the CellTiter- Glo Luminescent Cell Viability Assay kit. RESULT MLKL forms cation channels that are permeable preferentially to Mg2+ rather than Ca2+ in the presence of Na+ and K+. Moreover,each MLKL monomer contains five transmembrane helices:H1, H2, H3 , H5 and H6 of the N-terminal domain which is sufficient to form channels. Finally, MLKL-induced membrane depolarization and cell death exhibit a positive correlation to its channel activity.

Objective To survey knowledge of the local people's understanding of Brucellosis,and to explore the risk factors for brucellosis infection,and to determine the key issue of next comprehensive health promotion intervention.Methods Two counties,Yanggao and Guangling,which are old endemic areas with Brucellosis in history,and with epidemic rebounding in recent years,were selected.The survey was carried out by two stage stratified cluster sampling method.The questionnaires included respondents' demographic data(gender,age,education level,etc.),Brucellosis (hereinafter referred to as Brucellosis) knowledge of the investigation and behavior and attitude of people toward the measure for control of Brucellosis.Results A total of 5372 people were investigated in two counties of which 62.7％(3362/5372) of farmers.The investigated crowd had low culture level.The awareness of Brucellosis infection route of Yanggao and Guangling counties were 84.03％ (2379/2831) and 333％(847/2541 ).The average awareness of Brucellosis infection route was 18.60％(6001/32 260).In the investigation of knowledge on Brucellosis prevention of the two counties,29％ believed that it was necessary to wear gloves to process flow product,and 70％ of people answered do not know.For farmers on how to deal with dead animals,results showed that 79.1％(664/839) in Yanggao choose to sell dead animals to the market; 61.2％ (267/361) in Guangling choose to kill and bury,there were inappropriate treatment on handling of ill and dead animals in the two counties.Conclusions Spread of Brucellosis is caused mainly due to emphasis on the disease is not enough,and inappropriate handling of dead livestock.Measures like strengthening health education and behavioral intervention,increasing public awareness of the disease prevention and ability to change the incorrect way of life and cognitive concepts,can effectively reduce human infection and the spread of Brucellosis.

?AlM: To investigate the outcome and safety of Ahmed glaucoma valve implantation treatment in uncontrolled primary congenital glaucoma ( PCG) .
? METHODS: Twenty - two eyes in 22 children with uncontrolled PCG were reviewed retrospectively and underwent Ahmed glaucoma valve implantation treatment from January 2011 to December 2014. Main checking index included intraocular pressure ( lOP ) before and after operation, corneal diameter and complications.
?RESULTS: Preoperative mean age was 3. 74±2. 24y, and 2. 59 ± 1. 78y apart from the last operation. Postoperative average lOP was 35. 22 ± 6. 36mmHg. Average corneal diameter was 12. 79 ± 0. 75mm. Mitomycin C ( 0. 3 - 0. 5mg/mL ) was used in all operations for 3-5min. Glaucoma valves were implanted in the temporal or nose above the equator sclera. Postoperative lOP was 11. 4±4. 45mmHg at 1wk, and 16. 73± 7. 23mmHg after 12mo. As lOP< 21mmHg for success criteria, lOP of 16 eyes ( 73%) were controlled after 12mo. Preoperative 6 cases had shallow anterior chamber, recovered spontaneously. No serious complication was recorded, such as rejection of glaucoma valve, endoophthalmitis and corneal decompensation.
?CONCLUSlON:Ahmed glaucoma valve implantation in uncontrolled PCG is a safe and viable treatment.

Objective:To investigate the feasibility of targeted imaging and therapy of prostate cancer using nanocomposite probes composed of fluorescent magnetic nanoparticles(FMCNPs) and single chain Fv(ScFv) antibody specific for gama-seminoprotein.Methods:The nanocomposite probes(FMCNPs-ScFv) were prepared by conjugating fluorescent magnetic nanoparticles with singlegama-chain Fv antibody specific gama-seminoprotein,and were characterized by high resolution transmission electron microscopy,fluorescent spectrum and magnetic spectrum.Nanocomposite probes were incubated with prostate cancer LNCaP cells,and the targeting results of nanocomposite probes were observed by fluorescent microscopy.The cytotoxicity effect of the nanocomposite probes was measured by MTT.Nude mice models of prostate cancer were established and identified by immunohistochemistry method.The nanocomposite probes were injected into nude mice via tail vein.The distribution of nanocomposite probes in the nude mice was observed by Micro-animal imaging system,targeted imaging of the prostate cancer was observed by MR instrument.The nude mice with prostate cancer were irradiated with 100 W magnetic field for 30 min,and the changes of tumor sizes were observed.Results:The FMCNPs-ScFv nanocomposite probes were successfully prepared.Nanocomposite probes entered into the cytoplasm of cancer cells and exhibited low cytotoxicity effect.Nude mice model with prostate cancer were successfully fabricated;the nanocomposite probes distributed quickly in the main organs of mice,and gradually concentrated on the tumor tissues within 24 h.MR images showed that the tumor images were gradually enhanced from 6 h to 24 h after injection of the nanocomposite probe.Four days after magnetic irradiation,the tumors in the nude mice grew slower compared with the control nude mice(P