CHANGES OF MICROTUBULAR ORGANIZATION DURING THE EMBRYO SAC DEVELOPMENT IN RICE

Changes in the patterns of microtubular distributions and organization in various stages of development of the embryo sac in rice ( Oryza saliva L.) were different. In the megasporocyte, most microtubules were radially distributed but some were longitudinally oriented. Similar distributional patterns were seen in the dyad cells and the functional megaspore. Microtubules in the uni-nucleate embryo sac were mostly randomly distributed, although some radiated type was also present. The pattern of distribution of the microtubules in the two and four-nucleate embryo sac was quite similar and the microtubules were mainly of the perinuclear type. Microtubules in the cells of the eight-nucleate embryo sac were more complex. In the egg cell, the microtubules were mostly randomly distributed whereas in the synergids they were predominantly in longitudinal alignment. Those in the central cell were transversely aligned. The antipodal mass had very few microtubules and few longitudinally aligned ones were present in the cytoplasm.

外观单侧波形橡胶防护套的研制

介绍线缆用方形耐屈挠护套的配方设计、产品结构设计、生产工艺及产品性能特点。产品的径向截面设计成磨角长方形 ,外表面设计成单侧波形 ,因此 ,该产品表面美观大方、耐屈挠性好、使用寿命长 ,同时 ,由于表面积增大使得护套具有良好的耐磨性能。半成品设计成径向截面不封闭的管胚 ,这样具有硫化时装模方便、易于分段连续硫化等优点。

聚丙烯输液瓶吹塑模具设计

聚丙烯输液瓶是一种常用的大输液包装容器,其模具属于吹塑模具中的一种,设计吹塑模具时需要考虑吹塑模具的分型面、排气槽和冷却水等细节。现对该容器进行工艺分析,并从结构设计、排气槽设计、冷却3方面对其模具的吹制进行具体介绍。

提高胶管挤出质量措施的研究

主要阐述了我国橡胶挤出生产的现状，介绍了胶管挤出工艺和生产设备，分析管胚挤出中常见的质量问题，提出了保障管胚挤出质量的措施，以供探讨和借鉴。

Notch signaling dependent differentiation of cholangiocyte-like cells from rhesus monkey embryonic stem cells

Rhesus monkey embryonic stem（rES） cells have similar characteristics to human ES cells,and might be useful as a substitute model for preclinical research.Notch signaling is involved in the formation of bile ducts,which are composed of cholangiocytes.However,little is known about the role of Notch signaling in cholangiocytic commitment of ES cells.We analyzed the effect of Notch signaling on the induction of cholangiocyte-like cells from rES cells.About 80% of definitive endoderm（DE） cells were generated from rES cells after treatment with activin A.After treatment with BMP4 and FGF1 on matrigel coated wells in serum-free medium,rES-derived DE gave rise to cholangiocyte-like cells by expression of cholangiocytic specific proteins（CK7,CK18,CK19,CK20,and OV-6） and genes（GSTPi,IB4,and HNF1β）.At the same time,expression of Notch 1 and Notch 2 mRNA were detected during cell differentiation,as well as their downstream target genes such as Hes 1 and Hes 5.Inhibition of the Notch signal pathway by L-685458 resulted in decreased expression of Notch and their downstream genes.In addition,the proportion of cholangiocyte-like cells declined from ～90% to ～20%.These results suggest that Notch signaling may play a critical role in cholangiocytic development from ES cells.

Further Studies on Microtubule Organizational Changes During Megagametogenesis in Rice Embryo Sac

Changes in the pattern of microtubule distribution and organization during megagametogenesis in the embryo sac of rice (Oryza sativa L. cv. IR36) were re-examined using a modified polyethylene glycol sectioning technique before immuno-fluorescence staining of microtubules. In the sectioned materials the pattern of distribution and structural organization of the microtubule cytoskeleton were quite well preserved. Fine details of the patterns of structural changes and re-organization of the microtubule cytoskeleton in the major stages of development during embryo sac megagametogenesis (viz. functional megaspore, uni-nucleate, 2-nucleate, 4-nucleate, 8-nucleate and mature stage) could be clearly observed and easily followed. Some new organizational patterns of microtubules associated with the probable movement and positioning of the polar nuclei were observed.

Tissue Culture and Rapid Multiplication Techniques of Apocynum L.

[Objective] This study aimed to investigate rapid multiplication of Apocynum by tissue culture so as to provide plantlet sources for its industrialized cultivation. [Method] The asepsis seedlings were obtained by dealing with Apocynum seeds. Its cotyledons, hypocotyls and shoot tips were cultured on the media containing different concentrations of hormones. Finally, the influence of different hormone combinations on differentiation of cotyledons and hypocotyls, rapid multiplication of shoot tips, rapid multiplication of regenerated shoots, and rooting of test-tube plantlets was com- pared. [Result] MS＋2.0 mg/L BA＋0.03 mg/L NAA and MS＋0.07 mg/L NAA were the optimum medium for inducing regenerated buds from cotyledons and hypocotyls re- spectively; MS＋2.0 mg/L BA＋0.02 mg/L NAA was the best medium for rapid multi- plication of shoot tips; MS＋1.9 mg/L BA＋I.7 mg/L NAA was the best medium for rapid multiplication of regenerated buds： and 1/2MS＋0.6 mg/L NAA was the best medium for inducing roots. [Conclusion] The optimum hormone combination was de- termined for Apocynum rapid multiplication by tissue culture, which provides technical support on Apocynum industrialized cultivation.

Tissue engineering of blood vessels with endothelial cells differentiated from mouse embryonic stem cells

Endothelial cells (TEC_3 cells) derived from mouse embryonic stem (ES) cells were used as seed cells to construct blood vessels. Tissue engineered blood vessels were made by seeding 8 × 10~6 smooth muscle cells (SMCs) obtained from rabbit arteries onto a sheet of nonwoven polyglycolic acid (PGA) fibers, which was used as a biodegradable polymer scaffold. After being cultured in DMEM medium for 7 days in vitro, SMCs grew well on the PGA fibers, and the cell-PGA sheet was then wrapped around a silicon tube, and implanted subcutaneously into nude mice. After 6～8 weeks, the silicon tube was replaced with another silicon tube in smaller diameter, and then the TEC_3 cells (endothelial cells differentiated from mouse ES cells) were injected inside the engineered vessel tube as the test group. In the control group only culture medium was injected. Five days later, the engineered vessels were harvested for gross observation, histological and immunohistochemical analysis. The preliminary results demonstrated that the SMC-PGA construct could form a tubular structure in 6～8 weeks and PGA fibers were completely degraded. Histological and immunohistochemical analysis of the newly formed tissue revealed a typical blood vessel structure, including a lining of endothelial cells (ECs) on the lumimal surface and the presence of SMC and collagen in the wall. No EC lining was found in the tubes of control group. Therefore, the ECs differentiated from mouse ES cells can serve as seed cells for endothelium lining in tissue engineered blood vessels.

Accessory Breast Cancer Occurring Concurrently with Bilateral Primary Invasive Breast Carcinomas:A Report of Two Cases and Literature Review

The development of accessory breast tissue,which is found anywhere along the milk line,is attributed to the failure of milk line remnants to regress during embryogenesis.Primary tumors may arise from any ectopic breast tissue.Accessory breast cancer occurring concurrently with primary invasive breast cancer is extremely rare.Two such cases were reported in this article.One was a 43-year-old Chinese female who exhibited bilateral breast cancer(invasive ductal carcinoma,not otherwise specified,IDC-NOS) and an accessory breast carcinoma(IDC-NOS) incidentally identified in her left axilla.The ectopic breast tissue in her right axilla presented with adenosis.The patient was surgically treated,followed by postoperative docetaxel epirubicin(TE) chemotherapy.The second case was a 53-year-old Chinese female with bilateral breast cancer(apocrine carcinoma) accompanied by an accessory breast carcinoma(IDC-NOS) in her right axilla that was also incidentally identified.The patient was surgically treated after three doses of cyclophosphamide epirubicin docetaxel(CET) neoadjuvant chemotherapy,followed by adjuvant chemotherapy of the same regimen.

Clinicopathological and prognostic analysis of 429 patients with intrahepatic cholangiocarcinoma

AIM:To understand the clinicopathological characteristics and treatment selections and improve survival and provide valuable information for patients with intrahepatic cholangiocarcinoma(ICC). METHODS:We retrospectively evaluated 5311 liver cancer patients who received resection between October 1999 and December 2003.Of these,429(8.1%)patients were diagnosed with ICC,and their clinicopathological, surgical,and survival characteristics were analyzed. RESULTS:Upper abdominal discomfort or pain(65.0%), no symptoms(12.1%),and hypodynamia(8.2%)were the major causes for medical attention.Laboratory tests showed 198(46.4%)patients were HBsAg positive, 90(21.3%)hadα-fetoprotein>20μg/L,50(11.9%) carcinoembryonic antigen>10μg/L,and 242(57.5%) carbohydrate antigen 19-9(CA19-9)>37 U/mL.Survival data was available for 329(76.7%)patients and their mean survival time was 12.4 mo.The overall survival of the patients with R0,R1 resection and punching exploration were 18.3,6.6 and 5.6 mo,respectively. Additionally,CA19-9>37 U/mL was associated with lymph node metastases,but inversely associated withcirrhosis.Multivariate analysis indicated that radical resection,lymph node metastases,macroscopic tumor thrombi and size,and CA19-9 were associated with prognosis. CONCLUSION:Surgical radical resection is still the most effective means to cure ICC.Certain laboratory tests(such as CA19-9)can effectively predict the survival of the patients with ICC.

Agrobacterium-mediated transformation of herbicide resistance in creeping bentgrass and colonial bentgrass

Embryogenic calli were induced from the seeds of creeping bentgrass ( Agrostis palustris Huds.) cv. Regent and colonial bentgrass ( Agrostis Tenuis Sibth. Fl. Oxen.) cv. Tiger. The embryogenic calli were precultured on fresh medium for 4-7 days and then co cultivated with Agrobacterium tumefaciens , LBA4404, which contains plasmid vector pSBGM harboring bar coding region, synthetic green fluorescent protein (sGFP) coding region and matrix attachment region (MAR). After 3 days of co cultivation, the calli were washed thoroughly and transferred to MS medium containing 2 mg/L of 2, 4 D, 12-15 mg/L phosphinothricin (PPT) and 250 mg/L of cefotaxime. After 2-3 months of selection, the actively growing calli of 'Regent' and 'Tiger' were transferred to MS medium with 12-15 mg/L PPT and 250 mg/L cefotaxime for regeneration. The putative transformants were maintained on MS medium with 3 mg/L PPT for long period but control died within 1 month. After establishing in greenhouse, the transformants also showed strong resistance to 0.4% of herbicide Basta but control plants died within 2 weeks. Under confocal microscope, both young leaves and roots showed significant GFP expression. PCR analysis revealed the presence of a DNA fragment of GFP gene at the expected size (380 bp) in the transformants and its absence in a randomly selected control plant.

Studies on the role of microtubules in myofibrillogenesis

Co-localization of mierotubule (MT) and muscle myosin (MHO) myofibril immunofluoresoence in developing myotubes of chicken' skeletal muscle cultures was observed by using double staining of tubulin and MHO indirect immunofluorescence. 12-o-tetradeeanoyl-phorbol-13-acetate (TPA) selectively and reversibly blocks mycfibrillogenesis and alters the morphology of myotubes into myosacs where MTs are present in radiating pattern,when the arrested myogenic cells recover and start myofibrillogenesis after released from TPA, prior to the emergence of myofibrils, the pre-existing MTs become bipolarly aligned ooincidently with the tubular restoration of cell shape. Single nascent myofibrils overlapping with MTs extend into the base of growth tips where MTs go farther to the end of the tips. That MT might act as scaffold in guiding the bipolar elongation of the growing myofibrils was suggested. Taxol and ooloemid disturbed MT polymerization and disposition, and interfered with the normal spatial assembly of myofibrils in developing myotubes.

Antibodies against the C-terminal peptide of rabbit oviductin inhibit mouse early embryo development to pass 2-cell stage

A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductins. N-terminal peptide (NTP) (384 residues) and C-terminal peptide (CTP) (73 residues) of deduced protein precursor has about 80% and 50% identity with that of other mammals respectively. Fusion proteins GST-NTP 368(1R-368N)and GST-CTP73 (369F-441A) were expressed and purified. NH2-terminal of CTP sequencing reveals that the purified protein is consistent with the deduced one. In order to study the function of NTP and CTP the mouse anti-NTP and rabbit anti-CTP antisera were prepared. Tissue-specific (skeleton muscle, oviduct, uterus, ovary, liver, heart and brain) analysis indicated that rabbit oviductin was only found in oviduct. The conditioned medium derived from the rabbit oviduct mucosa epithelial cells has a function of overcoming the early embryonic development block of Kunming mous e cultured in vitro. Anti-CTP antiserum could totally inhibit the early embryo development at 2-cell stage cultured in the conditioned culture medium, but anti-NTP antiserum couldn’t. There was a positive relationship between the ratio of early embryos at development block and the dosage of anti-CTP antiserum added in the conditioned culture medium. These results suggest that oviductin has a function not only on fertilization, but also on the release of early embryonic development block, and the later function domain of rabbit oviductin may be situate in its C-terminal.

VEGFR-3 ligand-binding and kinase activity are required for lymphangiogenesis but not for angiogenesis

Although VEGFR-3 deficiency disrupts blood vascular development during early embryogenesis, the underlying mechanism was not clear. To characterize its function in angiogenesis and lymphangiogenesis, we employed two genetically modified mouse models in this study, targeting the coding region for the ligand-binding domain （Vegfr△LBD） or the tyrosine kinase domain with an inactivation point mutation （Vegfr3^TKmat）. We show that lymphatic growth was disrupted in Vegfr3△LBD/△LBD and Vegfr3^TKmut3^TKmat mice, but blood vessels developed normally in both embryo and yolk sac. Interestingly, in Vegfr3△LBD/△LBD but not Vegfr3^TKmut3^TKmat mice, lymph sac was present but there was lack of iym- phangiogenic sprouting. We further demonstrate that both the wild-type and mutant forms of VEGFR-3 could form heterodimers with VEGFR-2, and decreased the level of phospho-VEGFR-2 and the downstream phospho-Erk1/2 in endothelial cells when they were treated with VEGF-A. These findings indicate that signaling mediated via VEGFR-3 activation by its cognate ligands （VEGF-C/-D） is not required for angiogenesis, and that VEGFR-3 may play a role in this process by modulating VEGFR-2-mediated signals.

Effects of the Development of Thyroid and Sex Glands on the Synthesis of Brain Tubulin in Chick and Mouse Embryos

Using the specific affinity of tubulin for colchicine and the strong absorption of tubulin to DEAE ion exchangers at neutral pH and moderate ionic strength,the amounts of tubulin in the brain from both mice and chicks during different developing stages were measured by ~3H-colchicine assay (expressed as colchicine binding activity).The results show that the rate oftubulin synthesis reached a peak value during the critical period of brain development.This is exactly the period during which the organization and function of thyroid are being perfected.Besides,during breeding period,the difference of tubulin contents between male and female is significant(P<0.001).The synthesis of tubulin is strictly sex dependent(this phenomenon appeared only during sex maturation stage).It is suggested that sexual hormones might exert their effect on tubulin synthesis.

Involvement of XZFP36L1, an RNA-binding protein, in Xenopus neural development ＇

Xenopus ZFP36L1 （zinc finger protein 36, C3H type-like 1） belongs to the ZFP36 family of RNA-binding proteins, which contains two characteristic tandem CCCH-type zinc-finger domains. The ZFP36 proteins can bind AU-rich elements in 3＇ untranslated regions of target mRNAs and promote their turnover. However, the expression and role of ZFP36 genes during neural development in Xenopus embryos remains largely unknown. The present study showed that Xenopus ZFP36L1 was expressed at the dorsal part of the forebrain, forebrain-midbrain boundary, and midbrain-hindbrain boundary from late neurula stages to tadpole stages of embryonic development. Overexpression of XZFP36L1 in Xenopus embryos inhibited neural induction and differentiation, leading to severe neural tube defects. The function of XZP36L1 requires both its zinc finger and C terminal domains, which also affect its subcellular localization. These results suggest that XZFP36L1 is likely involved in neural development in Xenopus and might play an important role in post-transcriptional regulation.

GPCR-CARMA3-NF-kappaB Signaling Axis： A Novel Drug Target for Cancer Therapy

G protein-coupled receptors （GPCRs） play pivotal roles in regulating various cellular functions. It has been well established that GPCR activates NF-κB and aberrant regulation of GPCR-NF-κB signaling axis leads to cancers. However, how GPCRs induce NF-κB activation remains largely elusive. Recently, it has been shown that a novel scaffold protein, CARMA3, is indispensable in GPCR-induced NF-κB activation. In CARMA3-deficient mouse embryonic fibroblast cells, some GPCR ligand-, like lysophosphatidic acid （LPA）, induced NF-κB activation is completely abolished. Mechanistically, upon GPCR activation, CARMA3 is linked to the membrane by β-arrestin 2 and phosphorylated by some PKC isoform. Phosphorylation of CARMA3 unfolds its steric structure and recruits its downstream effectors, which in turn activate the IKK complex and NF-κB. Interestingly, GPCR （LPA）-CARMA3-NF-κB signaling axis also exists in ovarian cancer cells, and knockdown of CARMA3 results in attenuation of ovarian cancer migration and invasion, suggesting a novel target for cancer therapy. In this review, we summarize the biology of CARMA3, discuss the GPCR （LPA）-CARMA3-NF-κB signaling axis in ovarian cancer and speculate its potential role in other types of cancers. With a strongly increasing tendency to identify more LPA-like ligands, such as endothelin-1 and angiotensin II, which also activate NF-κB through CARMA3 and contribute to myriad diseases, GPCR-CARMA3-NF-κB signaling axis is emerging as a novel drug target for various types of cancer and other myriad diseases.

Inhibition of angiogenesis by DADAG in vivo and in vitro

1,2,5,6-Dianhydro-3,4-diacetyl-galactitol （DADAG）, an alkylating sugar alcohol derivative, has been shown effective against tumor growth. In this research, we explored the effect of DADAG on angiogenesis in chick chorioallantoic membrane （CAM） model and on the proliferation and migration of human umbilical vein endothelial cells （HUVECs）. We also studied the possible mechanism of the anti-angiogenesis effect of DADAG. The results showed that DADAG （100, 500 and 1000μnol/L） inhibited angiogenesis in CAM model dose-dependently. Sulforhodamine B （SRB） assay indicated that DADAG （45, 90, 135, 180 and 225 μmol/L） suppressed HUVECs proliferation in a dose-dependent and time-dependent manner. High Content Screening （HCS, Cellomics） assay, in which the influence of cell proliferation on migration could be excluded, indicated that DADAG （45, 135 and 225 ~xmol/L） directly inhibited the motility ofHUVECs. Immunofiuorescence assay suggested that DADAG inhibited angiogenesis possibly by decreasing vascular endothelial growth factor （VEGF） expression in HUVECs. Our findings reveal that DADAG show anti-angiogenic activity in vivo and in vitro, which is related to the downregulation of VEGF expression in endothelial cells.

A survey of acupuncture effect on the result of invitro fertilization

Objective This study was made to evaluate the effect of acupuncture on fertilization results in the patients undergoing medical treatment with IVF. Methods This study was conducted on 164 infertile patients who had referred to the infertility clinic of Imam Khomeini Hospital in 2009–2010（82 patients in acupuncture group and 82 in control group）. In the acupuncture group, before embryo transfer on oocyte puncture day, 2 days after puncture and one cycle before embryo transfer the patients were put under electroacupuncture on the Bǎihuì（百会 GV 20）, Yāoyángguān（腰阳关 CV 3）, Tàichōng（太冲 LR 3）, Nèiguān（内关 PC 6）, Sānyīnjiāo（三阴交 SP 6）, Guīlái（归来 ST 29）, Zúsānlǐ（足三里 ST 36）, and Hégǔ（合谷 LI 4） for three times each for 25 minutes. The points on ears include nèi Shēngzhíqì（内生殖器 TF 2）, and Shénmén（神门 TF 4）. The control group received no acupuncture. Positive pregnancy test, clinical pregnancy, ongoing pregnancy were compared between the two groups. Results Pregnancy level in the acupuncture group were 34 pregnancies（41.5%） as compared to 21（25.6%） in the control group（P〈0.05）.Observation level of gestational sacs in the 5th week of pregnancy included 31（37.8%） and 17（20.7%） sacs in the acupuncture and control groups, respectively（P〈0.05）.Level of ongoing pregnancies after 12 weeks included 21（21.6%） and 12（14.6%） pregnancies in the acupuncture and control groups, respectively（P〉0.05）. Conclusion Acupuncture will considerably improve the result of IVF as compared to the control group.

Function of FEZF1 during early neural differentiation of human embryonic stem cells

The understanding of the mechanism underlying human neural development has been hampered due to lack of a cellular system and complicated ethical issues. Human embryonic stem cells (hESCs) provide an invaluable model for dissecting human development because of unlimited self-renewal and the capacity to differentiate into nearly all cell types in the human body. In this study,using a chemical defined neural induction protocol and molecular profiling, we identified Fez family zinc finger 1 (FEZF1) as a potential regulator of early human neural development. FEZF1 is rapidly up-regulated during neural differentiation in hESCs and expressed before PAX6, a well-established marker of early human neural induction. We generated FEZF1-knockout H1 hESC lines using CRISPR-CAS9 technology and found that depletion of FEZF1 abrogates neural differentiation of hESCs. Moreover,loss of FEZF1 impairs the pluripotency exit of hESCs during neural specification, which partially explains the neural induction defect caused by FEZF1 deletion. However, enforced expression of FEZF1 itself fails to drive neural differentiation in hESCs,suggesting that FEZF1 is necessary but not sufficient for neural differentiation from hESCs. Taken together, our findings identify one of the earliest regulators expressed upon neural induction and provide insight into early neural development in human.