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Chronic pain (CP) adversely impacts people’s health. However, the influence of CP on pregnant women and their fetuses remains unclear. Few reasonable interventions have been offered to these women. This study aimed to examine the effects of enriched environmental intervention (EEI) on the delivery outcome and offspring health of pregnant mice with CP. Forty pregnant mice were equally and randomly divided into four groups (control, sham, CP, and CP + EEI). Monoarthritis was induced by subcutaneous injection of the complete Freund’s adjuvant (100 μg/100 μl) into the left hind paw of mice on Day 0 of gestation. Sham mice received 100 µl of sterile phosphate-buffered saline subcutaneously. General health status and delivery outcomes of pregnant mice and general physical status, growth, and development of offspring were observed and compared between groups. CP mice exhibited less weight gain, higher rate of premature delivery, smaller litter size, lower live birth rate, lower filial birth weight, shorter filial tail length, and lower filial rates of achieving righting reflex and crawling compared to mice in the control/sham/CP + EEI groups. There were no significant differences between mice in the CP + EEI group and those in the control and sham groups. CP during pregnancy led to serious adverse impacts on both the mother and fetus mice in this study, and EEI was an effective treatment for these effects.

Vinorelbine (VIN) is a semi-synthetic vinca alkaloid and is one of the most active agents for the treatment of solid tumors. The drug is commonly administered through a peripheral vein. Although VIN is known to cause local venous toxicity, such as drug-induced phlebitis, the mechanism responsible for the toxicity remains unclear. To investigate the role of Toll-like receptor 4 (TLR4) in VIN-induced vascular endothelial injury, human umbilical vein endothelial cells (HUVECs) were prepared from umbilical cords recovered with the written informed consent of the parents and treated with VIN for 60 min. Following the washing away of the VIN, the cells were cultured for a further 6 and 12 h, and the changes in TLR4 expression in the HUVECs were detected by quantitative polymerase chain reaction and western blot analyses. Finally, the effects of VIN on the translocation of nuclear factor κB (NF-κB) were determined by immunofluorescence and confocal analysis. The VIN-treated cells were stretched, extended and irregular while the control cells exhibited normal morphology. The TLR4 mRNA and protein levels were significantly higher in the VIN-treated HUVECs than those in the control group (P

采用问卷调查结合访谈的方式，调查中国境内 冀东地区一个农村社区的语言文化生活和社区内 语言类传统民间艺术的活力。结果显示，调查地主 要以当地汉语方言和汉语普通话为交际语，二者既 互补又存在竞争。当地汉语方言是社区成员的母语 和主要交际语，汉语普通话仅是部分被调查对象在 特定交际场合使用的语言变体。社区成员对当地汉 语方言和汉语普通话的认同度都比较高，但二者侧 重点不同。对当地汉语方言的认同主要体现在情感 方面，认为当地汉语方言好听、亲切；对普通话的 认同表现为工具性，认为普通话有用、有助于找到 理想的工作。电视节目和现代音乐是调查地主要的 文化消费品，这些产品几乎全部采用普通话为载 体。由这些信息可以看出，虽然当地汉语方言是调 查地的主要语言，但汉语普通话越来越被社区成员 接受，其社会功能正在逐渐扩大。在这样的语言环 境下，当地传统的语言类民间艺术的生存空间越来 越小，仅15.6%的被调查对象自报经常收听或收看 传统戏曲，仅13.3%的被调查对象自报能够表演当 地语言类传统民间艺术，并且这些被调查对象多为 老年被调查对象。照此趋势发展下去，当地语言类 传统民间艺术的传承和发展令人担忧。基于以上事 实，本研究尝试提出了保护和发展当地传统民间艺 术的策略。 The current research studies the language and culture life in a rural community in Jidong area with the investigation methods of questionnaire and interview. It also analyses the vitality of traditional folk arts in the community. The investigation results show that the rural community is a diglossia community with the local Chinese dialect (LCD) and the Modern Standard Chinese (MSC) as its communicative tools. The two varieties complement and compete with each other. The former is the mother tongue and the main communicative language of the community members while the latter is a language variant used by a few of the respondents in some specific communication situations. Community members hold positive attitudes towards the two language varieties but from different perspectives. They view the LCD as a nice and cordial variety from emotional perspective, while the MSC as a useful language variety from instrumental perspective. TV program and modern music are the main cultural consumption goods in the community, which are performed by the MSC. The aforementioned information indicates that there is a trend that the MSC is replacing the LCD as the daily communication tool in the community with the MSC expanding its social functions. In this language circumstance, the social space of the local folk arts related to language is getting narrower and narrower. The investigation results show that only 15.6% of the respondents often listen to or watch traditional opera, while only 13.3% can perform the local traditional opera; In addition, most of those who listen to, watch or perform the local traditional opera are the elder people. The aforementioned findings indicate that the local folk arts are in the way of getting endangered. At the last part the study attempts to put forward some strategies of protecting and the local traditional folk arts.

The Notch signaling pathway is critical for the differentiation of many tissues and organs in the embryo. To study the consequences of Notch1 gain-of-function signaling on female reproductive tract development, we used a cre-loxP strategy and Amhr2-cre transgene to generate mice with conditionally activated Notch1 (Rosa(Notch1)). The Amhr2-cre transgene is expressed in the mesenchyme of developing female reproductive tract and in granulosa cells in the ovary. Double transgenic Amhr2-cre, Rosa(Notch1) females were infertile, whereas control Rosa(Notch1) mice had normal fertility. All female reproductive organs in mutants showed hemorrhaging of blood vessels progressing with age. The mutant oviducts did not develop coiling, and were instead looped around the ovary. There were multiple blockages in the lumen along the oviduct length, creating a barrier for sperm or oocyte passage. Mutant females demonstrated inflamed uteri with increased vascularization and an influx of inflammatory cells. Additionally, older females developed ovarian, oviductal, and uterine cysts. The significant change in gene expression was detected in the mutant oviduct expression of Wnt4, essential for female reproductive tract development. Similar oviductal phenotypes have been detected previously in mice with activated Smo and in beta-catenin, Wnt4, Wnt7a, and Dicer conditional knockouts, indicating a common regulatory pathway disrupted by these genetic abnormalities.

Wolong Nature Reserve is the largest reserve for protecting the endangered wild giant panda. Due to historical factors, even in many sections of the core protection area, the forests have been seriously destroyed and natural forests are poorly restored. However, the relative importance of the determinants for recruitment of communities under disturbance is rarely explored. In our study the endogenous and exterior factors in a forest gap that affect the conifer-broad-leaved mixed forest regeneration were investigated near Wuyipeng, one of the observation stations at Wolong, to explore which determinant had the greatest effect on gap regeneration and to discover the recruitment of seedling establishment in forest gaps. With a linear sampling method, environmental factors, gap characteristics and recruitment of new individuals were measured and examined in every forest gap along three sampling lines. Data of environmental factors in the gaps were collected for a Pearson correlation analysis in order to explore the disturbance and preprocessed characteristics of the gaps, using principal component analysis in SPSS. Correlation analysis was applied to further explore the relationship between changes in the gaps and the response of the regenerating seedlings. The results show that a range of natural and human disturbances affected the pattern and characteristics of the forest gaps in this area. The richness in the composition of the seedlings was higher than that of gap makers, but the order of dominance of the composition was different between seedlings and gap makers. The success of dominant species in establishing themselves was affected by different environmental factors. For instance, the establishment of Betula spp. was correlated significantly with topographic factors, while that of Abies faxoniana was affected by soil characteristics and that of Rhododendron spp. correlated significantly with topographic factors and characteristics of gap makers. Moreover, all the biodiversity indices of regenerating seedlings were significantly correlated with environmental principle components mostly reflecting soil properties. From this we can infer that soil characteristics are the most important factors affecting the regeneration of dominant species and seedling diversity under gap disturbances.

Small cell carcinoma of the cervix (SCCC) belongs to the neuroendocrine carcinomas, and it is a rare gynecological tumor of high-potential malignancy. It has a poorer prognosis compared to cervical squamous cancer or adenocarcinoma, and the therapeutic regimen of the disease differs. Diagnosis is based on pathomorphological characteristics, i.e., the small and round cancer cells (oat cell) which are uniform in shape and size, with the immunohistochemical marker helpful for diagnosis. Combined therapy is first recommended. Postoperative chemotherapy with platinum/etoposide (PE), vincristine/adriamycin/cyclophosphamide (VAC) and taxel/carboplatin (TP) can markedly improve the prognosis of early SCCC patients.

The Notch signaling pathway is critical for the differentiation of many tissues and organs in the embryo. To study the consequences of Notch1 gain-of-function signaling on female reproductive tract development, we used a cre-loxP strategy and Amhr2-cre transgene to generate mice with conditionally activated Notch1 (Rosa(Notch1)). The Amhr2-cre transgene is expressed in the mesenchyme of developing female reproductive tract and in granulosa cells in the ovary. Double transgenic Amhr2-cre, Rosa(Notch1) females were infertile, whereas control Rosa(Notch1) mice had normal fertility. All female reproductive organs in mutants showed hemorrhaging of blood vessels progressing with age. The mutant oviducts did not develop coiling, and were instead looped around the ovary. There were multiple blockages in the lumen along the oviduct length, creating a barrier for sperm or oocyte passage. Mutant females demonstrated inflamed uteri with increased vascularization and an influx of inflammatory cells. Additionally, older females developed ovarian, oviductal, and uterine cysts. The significant change in gene expression was detected in the mutant oviduct expression of Wnt4, essential for female reproductive tract development. Similar oviductal phenotypes have been detected previously in mice with activated Smo and in beta-catenin, Wnt4, Wnt7a, and Dicer conditional knockouts, indicating a common regulatory pathway disrupted by these genetic abnormalities.

Understory vegetation is an important habitat component for many wildlife species. Previous broad-scale studies on biodiversity and wildlife habitat have suffered from a lack of detailed information about understory distribution. Consequently, it is unclear how understory distribution influences the analysis of habitat quantity and spatial distribution. To address this problem, we compared estimates of giant panda (Ailuropoda melanoleuca) habitat with and without understory bamboo. The results show that the spatial distribution of bamboo has a substantial effect on the quantity and spatial arrangement of panda habitat. Total amount of estimated habitat decreased by 29-52% and decreased connectivity was notable after bamboo information was incorporated into the analyses. The decreases in the quantity and quality of panda habitat resulted in a decrease of 41% in the estimated carrying capacity. Our results suggest that it is necessary to incorporate understory vegetation into large-scale wildlife habitat research and management to avoid overestimation of habitat and improve broad-scale analyses of species distributions and biodiversity estimates in general. (C) 2004 Elsevier Ltd. All rights reserved.

To address the complex interactions between humans and wildlife habitat, we developed a concep- tual framework that links human factors with forested landscapes and wildlife habitat. All the components in the framework are integrated into systems models that analyze the effects of human factors and project how wildlife habitat would change under different policy scenarios. As a case study, we applied this frame- work to the Wolong Nature Reserve in Sichuan Province (southwestern China), the largest home of the giant panda ( Ailuropoda melanoleuca ). We collected ecological and socioeconomic data with a combination of var- ious methods ( field observations, aerial photographs, government documents and statistics, interviews, and household surveys) and employed geographic information systems and systems modeling to analyze and in- tegrate the data sources. Human population size has increased by 66% and the number of households in the reserve has increased by 115% since 1975, when the reserve was established. During the same period, the quality and quantity of the giant panda habitat dramatically decreased because of increasing human activi- ties such as fuelwood collection. Systems modeling predicted that under the status quo, human population in the reserve would continue to grow and cause more destruction of the remaining panda habitat, whereas re- ducing human birth rates and increasing human emigration rates would lower human population size and alleviate human impacts on the panda habitat. Furthermore, our simulations and surveys suggested that pol- icies encouraging the emigration of young people would be more effective and feasible than relocating older people in reducing human population size and conserving giant panda habitat in the reserve.

Previously we showed that a white pine protein Pin m III (a member of PR10 family of pathogenesis-related proteins) is up-regulated by infection in the white pine blister rust pathosystem. In this study, a blister rust fungal protein, Cro r I, which is similar in size to Pin m III (19 kDa), was detected in the infected white pine tissues. The N-terminal amino acid sequence of Cro r I isolated from infected pine foliage and from fungal mycelia was identical. Rabbit antibody was prepared to a synthetic N-terminal peptide and was purified by immunoaffinity. The purified antibody was used in a Western immunoblot to quantify the amount of Cro r I in various tissues. In western white pine seedlings the amount of Cro r I was significantly (p < 0.0001) higher in infected tissues of cankered seedlings than the infected tissues of resistant seedlings. In sugar pine seedlings, the amount of Cro r I was also significantly (p < 0.01) higher in infected tissues of susceptible seedlings than in resistant seedlings. Furthermore, Cro r I is secreted by the blister rust fungus and was found to be translocated to the healthy tissues of cankered white pines. Cro r I is a major protein that could be extracted from infected foliage by vacuum infiltration. The level of Cro r I detected in the mycelium of different isolates varied. The cDNA of Cro r I was isolated by reverse transcription - polymerase chain reaction. Comparison of the DNA sequence and the deduced protein sequence with data bases revealed that it is a previously undescribed protein. The calculated molecular weight from the deduced protein sequence of Cro r I was 16.7 kDa and the calculated isoelectric point was 9.55. Protein sequence analysis showed that Cro r I has two potential N-linked glycosylation sites in its sequence.Key words: translocation, elicitor, antibody, amino acid sequence.

Human population size and growth have been recognized as important factors affecting biodiversity, but the impacts of population structural changes on biodiversity are not clear. In this paper, we made the first attempt to link human population structural changes with implications for biodiveristy, using Wolong Nature Reserve (southwestern China) for the endangered giant panda as a case study. From 1982 to 1996, the labor force (20–59 years of age) in the reserve jumped by 59.76 percent, although the total population size increased by only 14.65 percent. During the same time period, the sex ratio (males:females) of small children (0–4 years of age) changed from 0.98:1 to 1.20:1, and the percentage of children receiving education beyond the elementary school level increased from 14.04 to 27.47. The increase in labor force and the number of male-biased children could have more negative impacts on the panda habitat, whereas improving school education could help more young people move out of the reserve by going to college and finding jobs elsewhere and thus reduce destruction to the panda habitat.

Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of crude extracts from the mycelium of five Beauveria bassiana isolates contained several proteins which were used to produce antibodies in BALB/c mice. The antibody response to these proteins was good and enzyme-linked immunoassay showed a wide antigenic cross-reactivity among the isolates. An isolate from Canada and one from the United States were the most cross-reactive while two isolates from China were the least cross-reactive. Both SDS-PAGE and Western immunoblot showed that each of the five isolates had a unique protein and antigenic profile. The immunoblot analyses also revealed the presence of a unique antigen in one of the Chinese isolates.

Calmodulin-dependent cyclic nucleotide phosphodiesterase is one of the key enzymes involved in the complex interactions, which occur between the cyclic nucleotide and Ca 2 + second-messenger systems. In eye, cAMP regulation is important in a variety of physiological processes such as aqueous humor regulation, photoreceptor signal transduction and retinal blood flow. Bovine eye calmodulin-dependent cyclic nucleotide phosphodiesterase was purified to apparent homogeneity and the isolated enzyme had a significantly higher affinity for calmodulin and Ca 2 + . Immunohistology revealed calmodulin-dependent cyclic nucleotide phosphodiesterase expression in corneal epithelium, retina and optic nerve of the eye. The cAMP-dependent protein kinase was found to catalyze the phosphorylation of bovine eye calmodulin-dependent cyclic nucleotide phosphodiesterase and the following observations were made. Firstly, the phosphorylation resulted in the incorporation of 1 mol of phosphate per mol of subunit, resulting in higher calmodulin and Ca 2 + concentration requirement for calmodulin-dependent cyclic nucleotide phosphodiesterase activation. Secondly, Ca 2 + and calmodulin prevented the phosphorylation. Thirdly, the phosphorylation of calmodulin-dependent cyclic nucleotide phosphodiesterase could be reversed by the calmodulin-dependent phosphatase, calcineurin. Analysis of the complex regulatory properties of the calmodulin-dependent cyclic nucleotide phosphodiesterase in the eye has led to the suggestion that fluxes of cAMP and Ca 2 + during cell activation are closely coupled and that calmodulin-dependent cyclic nucleotide phosphodiesterase plays a key role in this signal coupling phenomenon.

Calmodulin-dependent cyclic nucleotide phosphodiesterase is one of the key enzymes involved in the complex interactions, which occur between the cyclic nucleotide and Ca2+ second-messenger systems. In eye, cAMP regulation is important in a variety of physiological processes such as aqueous humor regulation, photoreceptor signal transduction and retinal blood flow. Bovine eye calmodulin-dependent cyclic nucleotide phosphodiesterase was purified to apparent homogeneity and the isolated enzyme had a significantly higher affinity for calmodulin and Ca2+. Immunohistology revealed calmodulin-dependent cyclic nucleotide phospho-diesterase expression in corneal epithelium, retina and optic nerve of the eye. The cAMP-dependent protein kinase was found to catalyze the phosphorylation of bovine eye calmodulin-dependent cyclic nucleotide phosphodiesterase and the following observations were made. Firstly, the phosphorylation resulted in the incorporation of 1 mol of phosphate per mol of subunit, resulting in higher calmodulin and Ca2+ concentration requirement for calmodulin-dependent cyclic nucleotide phosphodiesterase activation. Secondly, Ca2+ and calmodulin prevented the phosphorylation. Thirdly, the phosphorylation of calmodulin-dependent cyclic nucleotide phosphodiesterase could be reversed by the calmodulin-dependent phosphatase, calcineurin. Analysis of the complex regulatory properties of the calmodulin-dependent cyclic nucleotide phosphodiesterase in the eye has led to the suggestion that fluxes of cAMP and Ca2+ during cell activation are closely coupled and that calmodulin-dependent cyclic nucleotide phosphodiesterase plays a key role in this signal coupling phenomenon.

Calmodulin-dependent cyclic nucleotide phosphodiesterase was identified in and purified to apparent homogeneity from the total calmodulin-binding protein fraction of bovine eye in a single step by immunoaffinity chromatography. The bovine eye calmodulin-dependent cyclic nucleotide phosphodiesterase is immunologically similar to the bovine brain 60-kDa isozyme. The purified enzyme had higher affinity for calmodulin than the 60-kDa phosphodiesterase isozyme from bovine brain, but similar affinity to that of the heart isozyme. When the Ca2+-dependence of the eye enzyme was compared to cardiac calmodulin-dependent cyclic nucleotide phosphodiesterase at an identical concentration of calmodulin, the bovine eye calmodulin-dependent cyclic nucleotide phosphodiesterase was activated at the same Ca2+concentration as the bovine heart calmodulin-dependent cyclic nucleotide phosphodiesterase isozyme.

Calcineurin is a serine/threonine protein phosphatase which catalyzes the hydrolysis of both phosphoseryl/phosphothreonyl and phosphotyrosyl proteins as well as low molecular weight compounds such as p-nitrophenyl phosphate. It is a hetero-dimeric protein consisting of a 60 kDa A chain and 19 kDa B chain. Calcineurin A is organized into functionally distinct domains such as a catalytic domain, a calcineurin B binding domain, a calmodulin-binding domain, and an inhibitory domain. Calcineurin B has four EF-hand calcium binding domains with a secondary structure that is homologous to calmodulin but its metal binding properties are more similar to troponin-C. The N-terminal myristoyl group of calcineurin B might play a role in the interaction between subunits A and B during phosphorylation/dephosphorylation processes. Crystals of size 0.125 x 0.07 x 0.03 mm and 0.7 x 0.03 x 0.02 mm have been obtained for calcineurin and the A subunit respectively. Crystals of calcineurin show strong diffraction to 5.3 A and weak diffraction to 3.0 A on rotating anode operated at 50 kV and 100 mA. Further work is in progress to improve the X-ray diffraction quality of these crystals and to obtain well diffracting crystals of calcineurin B.

Calmodulin-dependent cyclic nucleotide phosphodiesterase (CaMPDE) is one of the key enzymes involved in the complex interactions that occur between the cyclic nucleotide and Ca2+ second messenger systems. This enzyme is characterized by a high Km for cAMP and a lower Km for cGMP. CaMPDE exists in different isozymic forms that exhibit distinct molecular and /or catalytic properties. Four CaMPDE isozymes have been purified close to homogeneity from bovine tissue in this laboratory. Immunologic, kinetic, and regulatory characterization have revealed subtle differences among these enzymes. The 63 kD CaMPDE is kinetically distinct from the other three isozymes. This isozyme is also further distinguished from other isozymes by the ginsenoside inhibition study. Ginsenosides are found to be potent inhibitors of brain 60 kD, heart and lung CaMPDE isozymes, but not of the brain 63 kD isozyme. The three other members — brain 60 kD CaMPDE, heart CaMPDE, and lung CaMPDE — show differential activation by calmodulin and Ca2+. These observations are consistent with the notion that differential regulation by calmodulin and Ca2+ kinetics inhibition are important functions of these isozymes that provide fine-tuning mechanisms for calmodulin action.

Calcineurin is a serine/threonine protein phosphatase which catalyzes the hydrolysis of both phosphoseryl/phosphothreonyl and phosphotyrosyl proteins as well as low molecular weight compounds such as p-nitrophenyl phosphate. It is a hetero-dimeric protein consisting of a 60 kDa A chain and 19 kDa B chain. Calcineurin A is organized into functionally distinct domains such as a catalytic domain, a calcineurin B binding domain, a calmodulin-binding domain, and an inhibitory domain. Calcineurin B has four EF-hand calcium binding domains with a secondary structure that is homologous to calmodulin but its metal binding properties are more similar to troponin-C. The N-terminal myristoyl group of calcineurin B might play a role in the interaction between subunits A and B during phosphorylation/dephosphorylation processes. Crystals of size 0.125 x 0.07 x 0.03 mm and 0.7 x 0.03 x 0.02 mm have been obtained for calcineurin and the A subunit respectively. Crystals of calcineurin show strong diffraction to 5.3 A and weak diffraction to 3.0 A on rotating anode operated at 50 kV and 100 mA. Further work is in progress to improve the X-ray diffraction quality of these crystals and to obtain well diffracting crystals of calcineurin B.

During male development, the testes move from a high intraabdominal position and descend into the scrotum. The gubernaculum, an inguinoscrotal ligament connecting the testis to the lower abdomen, is believed to play a critical role in this process. The first stage of testicular descent is controlled by insulin like3 hormone (INSL3), produced in testicular Leydig cells. Deletion of Insl3 or its receptor, Rxfp2, in mice causes cryptorchidism. We produced Cre/loxP regulated shRNA transgenic mice targeting RXFP2 expression. We have shown that the transgene was able to reduce Rxfp2 gene expression and thus behaved as a hypomorphic allele of Rxfp2. Variable degrees of uni- and bilateral cryptorchidism was detected in males with the activated shRNA transgene on an Rxfp2+/- background. Conditional suppression of Rxfp2 in the gubernaculum led to cryptorchidism. Gene expression analysis of a mutant cremasteric sac using Illumina microarrays indicated abnormal expression of a significant number of genes in Wnt/β-catenin and Notch pathways. We have demonstrated profound changes in the expression pattern of β-catenin, Notch1, desmin, and androgen receptor (AR), in Rxfp2-/- male embryos, indicating the role of INSL3 in proliferation, differentiation, and survival of specific cellular components of the gubernaculum. We have shown that INSL3/RXFP2 signaling is essential for myogenic differentiation and maintenance of AR-positive cells in the gubernaculum. Males with the deletion of β-catenin or Notch1 in the gubernacular ligament demonstrated abnormal development. Our data indicates that β-catenin and Notch pathways are potential targets of INSL3 signaling during gubernacular development.