Objective To observe the effects of endovascular radiation (ER) on the proliferation and apoptosis of medial smooth muscle cells (SMC) and to discuss the possible mechanisms of radiation in the prevention of vascular restenosis (RS) in rabbits after carotid endarterectomy (CEA).Methods Forty rabbits undergoing CEA were randomly divided into four groups (each group=10) and given a radiation dose of 0, 10, 20 and 40 Gy 32 P respectively. Rabbits were killed on the 3rd, 7th, 14th, 28th and 56th day after operation. The specimens were collected and histopathologic examinations were done.Results Proliferation apparently occurred in the intima and media of carotid the lumen became narrow in the control group on the 14 th, 28 th and 56 th day after operation. While in the radiation groups, proliferation was apparently suppressed and the lumen was much less narrowed ( P

Objective To construct the recombinant RIP3 over-expressed plasmids and transfect them in breast cancer MCF7 cells, and identify the expression and localization of fusion protein, as well as its effect on the death way of MCF7 cells. Methods The expression levels of RIP3 mRNA in four breast cancer cell lines and normal mammary epithelial cells were detected by reverse transcription polymerase chain reaction (RT-PCR). The RIP 3 coding sequence was amplified by polymerase chain reaction and subcloned into mCherry vector to construct recombinant plasmids. The plasmids were transfected into MCF7 cells by lentivirus after DNA sequencing, then screened by basticidin (4 mg/L) for 1 week. The efficiencies of RIP3 expression were validated by Western blotting assay. The death way of mCherry-RIP3-MCF7 cells was observed under the treatment of TNF-αand Z-VAD-FMK. Results The lowest expression of RIP3 mRNA was found in MCF7 cells. The sequencing results validated the well recombinant plasmids. The expression of mCherry-RIP3 fusion pro-tein with a molecular weight of 85 ku was detected by Western blot assay. The mCherry-RIP3 expression enhanced the sensi-tivity of MCF7 cells to TNF-αand Z-VAD-FMK induced cell death. Conclusion The recombinant RIP3 over-expressed plasmids were successfully constructed, and the stable MCF7 cells with ectopic RIP3 transfection were obtained. The mCher-ry-RIP3 fusion protein was expressed in the cytoplasm and was conformed to mediate TNF-αinduced necroptosis.

BACKGROUND:Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cels, but there is no report on the effect of autophagy, as wel as the interaction between autophagy and apoptosis. OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cels. METHODS:Density gradient centrifugation was used to obtain mononuclear cels from bone marrow, and the mononuclear cels were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cels colected were the endothelial progenitor cels. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cel cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cels in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-IIprotein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagywas particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cels was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cel apoptosis, change the cel cycle significantly, and can inhibit the proliferation of endothelial progenitor cels.

BACKGROUND:Human embryonic stem cels are able to self-renew indefinitely and have the capacity to differentiate into al three germ layers (ectoderm, endoderm and mesoderm). These properties imply great potential in the basic research and clinical application, including regenerative medicine, drug screening and toxins, early human embryo, cel transplantation, gene therapy,etc. However, it is a substantial chalenge to develop efficient techniques for their large-scale culture under defined conditions, and for controling and directing their differentiation. For therapeutic purposes, many scholars are trying to establish methods for maintaining pluripotency in defined xeno-free conditions and scalable culture systems. OBJECTIVE:To discuss the progress of serum-free culture systems in human embryonic stem cel research reported in recent years and to highlight the chalenges and advances being made towards the development of serum-free and xeno-free culture systems suitable for therapeutic applications. METHODS:A computer-based search of CNKI and PubMed academic database was performed for articles addressing serum-free culture systems of human embryonic stem cels published from 2008 to 2015. Repetitive and old articles were excluded. Finaly, 58 articles were summarized. RESULTS AND CONCLUSION:Several groups have attempted to exclude individual animal components by using feeder-free matrices, feeder cels of human origin, or defined xeno-free media, aiming to select a suitable matrix and medium that can minimize or not use heterologous components, in order to obtain cel lines at clinical level. However, the current cel products are far from clinical application. There are stil many problems to be solved, such as standardization, normalization and individualization of cel products. With the normative development of stem cel research and industry, human embryonic stem cel products are expected to be widely used in clinic.

From 6 stains of yeasts, Saccharomyces cerevisiae 2016 which was suitable for Cr rich yeasts fermentation was selected Concentration and additive method of CrCl 3 were investigated by liquid culture and the optimum conditions of Cr rich yeasts were obtained The optimum concentration of CrCl 3 were 5?10 4 mol/L that were added to liquid medium at the initial 8 h of culture time At this condition, the biomass of Cr rich yeasts was 1 55 g dry weight per 100 milliliter, Cr content in yeast cells was above 1100 ?g/g, the utilizable rate of Cr ion was about 60%

Objective To evaluate the strategies and effect of surgical treatment for concomitant abdominal aortic aneurysm (AAA) and colorectal cancer (CRC). Methods Literatures on concomitant AAA and CRC published from January 1988 to December 2008 were retrieved from Pubmed, Sciencedirect, Ovid, CBMdisc, CNKI and et al, and correlated indexes were extracted for analysis. Differences among the groups were analyzed using the t test, chi-square test and fisher's exact test. Results A total of 367 cases of concomitant AAA and CRC treated by operation were retrieved. The length of operation delay of patients who received radical resection of CRC first was (115 ± 21 )days, which was significantly longer than (42 ± 8 )days of patients who received open abdominal aortic aneurysm repair (OAAR) first (t = 18. 9, P <0.05). The 30-day complication rate and accumulative length of hospital stay of patients who received one-stage radical resection of CRC + OAAR were 10.5% ( 12/114 )and (23 ±6) days, and 26.0% (47/181) and ( 16 ±4)days of patients who received two-stage radical resection of CRC + OAAR, with a significant difference ( χ2 = 10.42, t = 12. 01, P <0.05 ). The accumulative length of hospital stay of patients who received radical resection of CRC + endovascular aneurysm repair (EVAR) was (12 ±4) days, which was significantly shorter than that of patients who received radical resection of CRC + OAAR [ ( 19 ±5 ) days ] ( t = 9.48, P < 0. 05 ). The 4-year survival rate of patients who received two-stage radical resection of CRC + OAAR was 43.5% (27/62), which was significantly lower than that of patients who received two-stage radical resection of CRC + EVAR [69.2% (18/26) ] or one-stage radical resection of CRC + OAAR [73.7%(14/19) ] (χ2 =4.83, 5.28, P<0.05). Conclusions If the diameter of AAA is under 5 cm, radical resection of CRC should be firstly carried out; but if the diameter of AAA is above 5 cm, OAAR should be firstly carried out to prevent the rapture of tumors. One-stage surgery is better than two-stage surgery if patients could tolerate it.

Objective To identify the genetic mechanism of fetuses with short limbs deformity.Methods From Aug.2008 to Aug.2011,ten fetuses with obvious short limbs were found in ultrasound screening performed at 18-24 and (or) 30-32 gestational weeks and underwent artificial induced labor with the patient' consent.Amniotic fluid or cord blood of the fetuses was collected for karyotyping analysis and detection of mutation point of fibroblast growth factor receptor 3 (FGFR3)gene by polymerase chain reaction and gene sequencing.One fetus (case 3) who presented with achondrogenesis underwent sequencing of SLC26A2 and Trip11 gene meanwhile.Results Among the 10 fetuses with short limbs deformity,five cases were found during second trimester and five during third trimester.Nine cases were identified as normal karyotype and one was chimera (46,XY/45,XY,- 18).One fetus carried a rare FGFR3 mutation of c.1108G＞T (G370C) and was diagnosed as thanatophoric dysplasia at 21+3 weeks.Three fetus carried c.1138G＞A (G380R) mutation and were diagnosed as achondroplasia.These four families had low recurrent risk because no gene mutations were found in the parents.Three mothers of these four fetuses were pregnant again and had normal neonates now.No mutations were found in all gene sequencing in case 3.Conclusions Karyotyping analysis and sequencing of FGFR3 gene could find causative gene mutations and provide genetic counselling and prenatal diagnosis for some fetuses with short limbs deformity.In the third trimester,achondroplasia is the most possible diagnosis when short limbs fetus is found by ultrasound.

Objective To establish the genetic test technique of trisomy 21 concurrently conducts with prenatal diagnosis for hereditary hearing loss. Methods Fifty-four pregnant women who underwent prenatal diagnosis for hearing loss of their fetuses in Chinese People's Liberation Army General Hospital from March 2009 to May 2010 were enrolled in this study. All probands from the deaf families have confirmed the causative mutation for hearing loss in Genetic Testing Center in Chinese People's Liberation Army General Hospital. The mean age of 54 pregnant women is 31 years at pregnancy of 18 - 26 weeks, 5 cases ＞ pregnancy of 23 weeks, 9 cases ≥ 35 years. All subjects did not conduct the serologic tests for trisomy 21before. Fifteen to twenty ml amniotic fluid was drawn from 49 cases at pregnancy of 18 - 23 weeks and 5 cases ＞ pregnancy of 23 weeks. One to two ml umbilical blood was drawn from 5 cases ＞ pregnancy of 23 weeks. For 9 cases ≥ 35 years, amniotic fluid cell culture and karyotyping analysis were conducted concurrently. A multiple quantitative fluorescent ( QF) PCR and six microsatellite markers were applied to as trisomy 21. Results (1) Fifty-four fetuses were successfully conducted prenatal genetic diagnosis for hearing loss (included GJB2 and SLC26A4). Ten fetuses copied the exactly same genotypes as the probands. The other 44 cases fetuses did not copy the same genotypes as the probands and won't develop hearing loss. The hearing test showed normal hearing for the neonates. (2) All the 54 fetuses were excluded of trisomy 21 by QF-PCR and were verified after birth. Five fetuses with advanced maternal age were performed karyotyping analysis and showed normal. The diagnostic results of QF-PCR can be obtained in 1 - 3 days without misdiagnosed. Conclusions QF-PCR is an efficient, rapid and accurate technique for detection of trisomy 21 without increasing sample amount. It can be used for fetuses who were undertaken hearing loss gene test or other prenatal gene test.

Objective To investigate the phenotype and genetic characteristics of a Chinese family with an autosomal-dominant inherited sensorineural hearing loss.Methods A Chinese pedigree associated with an autosomal-dominant inherited low-frequency sensorineural hearing loss (LFSNHL) was investigated.After obtaining informed consent from all study participants medical and audiological examination were used to rule out any syndromic hearing impairment.Five patients were tested with DPOAE and ABR,while two patients were tested with vestibular function and computed tomography scan of the temporal bone to exclude auditory neuropathy and other possible aural disorders.Twenty-one loci and twenty-three genes of DFNA screening had been done by using microsatellite markers and linkage analysis.Results Proband of the family had been diagnosed with low-frequency sensorineural hearing loss.A Chinese family BJ-L046 with non-syndromic autosomal dominant hearing loss was ascertained.Hearing impairment in the affected members in family BJ-L046 occured from 10 to 20 years of age and mainly affected the low frequencies,causing an upsloping audiogram.Higher frequencies were getting involved with increasing age,thus causing a flat-type audiogram.No positive result was found in twenty-one loci and twenty-three genes of DFNA screening.Conclusion A Chinese family with late-onset low-frequency sensorineural hearing loss was clinically studied.No positive result was found by linkage analysis,and twenty-one loci and twenty-three genes of DFNA were preliminary excluded.

Objective To explore the expression of V-ATPase in colon cancer and its clinical significance. Methods Detecting the expression of V-ATPase mRNA in 20 paired of colon tumor tissues and normal tissues by using reverse transcription-polymerase chain reaction ( RT-PCR) and real-time quantitative polymerase chain reaction( Real-time PCR) , and testing the expression of V-ATPase protein by immu-nohistochemistry of EnVinsion. Results The expression of V-ATPase mRNA in tumor tissues and its paired normal tissues were (5. 37 ± 0. 44) and (2. 03 ± 0. 35)(P<0. 01). The positive immunohistochemistry of V-ATPase in tumor tissues and its paired normal tissues were 69. 1%(47/68) and 5. 8%(4/68) respectively, and the positive expression were primarily in cytoplasm and cytomembrane. Overexpression of V-ATPase was associated with tumor stage (P<0. 05), lymph node metastasis (P=0. 044), distant metastasis (P=0. 049), vessel in-vasion (P=0. 044) and differentiation (P<0. 001). Conclusion Overexpression of V-ATPase plays a significant role in the carcinogene-sis and the progression of colon cancer, which might be an important postoperative therapeutic target.