苹果霉心病病原研究

１９８９－１９９３年，在甘肃苹果产区采集红星或元帅品种病果和无症状果实，逐果取果心组织分离，获得３０余个真菌分离物，以链格抱（Ａｌｔｅｒｎａｒｉａａｌｔｅｒｎａｔａ）出现率最高，占５１．３％；其次为粉红单端孢（Ｔｒｉｃｈｏｔｈｅｃｉｕｍｒｏｓｅｕｍ）、棒盘孢（Ｃｏｒｙｍｅｕｍｓｐ．）、节孢状镰刀菌（Ｆｕｓａｒｉｕｍａｒｔｈｒｏｓｐｏｒｉｏｉｄｅｓ）、狭截盘多毛孢（Ｔｒｕｎｃａｔｅｌｌａａｎｇｕｓｔａｔａ）等，依次占１１．８％、１２．０％、８．９％、７．９％。每个单果大多是只能分离出一种菌，少数出现２一３种菌。不同症状的果实，出现的真菌种类不同，霉心果以链格孢出现率最高，占６０％－８０％；心腐果中链格孢出现率显著较少，占１０％－３０％；而粉红单端孢、节孢状镰刀菌、棒盘孢、狭截盘多毛孢等５种菌的出现率较高，占７．８％－２５．５％。用果心不带菌的果实人工接种，对１２种分离物进行致病性测定看出，不同菌株之间致病力有明显差异，致心腐的病菌主要有５种，分别是粉红单端孢、棒盘孢、节孢状镰刀菌、狭截盘多毛孢和一种不产孢的浅色丝状菌。链格孢主要致霉心症状。混合接种试验表明，如有几种真菌进入果心，则致病性强的一种首先占据优势，?     

双孢蘑菇的单孢分离和单孢杂交研究

本文采用一种较为简单易行、高效、选择性强的单孢分离法，分离获得双孢蘑菇的单孢菌林１５８株，并对这些单孢菌株的培养性状、菌丝间融合和自发营养缺陷型等进行研究。结果表明：来自同一子实体的许多单孢子在ＰＤＡ培养基上其菌丝体形态、生长速度均有显著差异：单孢菌株之间的融合率为１８．１％；单孢菌株中存在自发营养缺陷型，自然发生率为１０．１５％；活蘑菇菌丝体及高浓度孢子悬液对于单孢子萌发均具明显的促进作用。     

应用菌刺洗脱法高效分离猴头菇单孢

单孢分离是食药用菌获得单核体菌株的重要手段之一,建立的菌刺洗脱法可对猴头菇进行简单高效的单孢分离。试验结果表明,这种方法直接将猴头菇菌刺上的担孢子洗下来,不需孢子收集器,操作简单、快速,能够有效降低污染率,缩短单孢分离的时间。     

枣缩果病原越冬场所调查

通过田间采集枣树的树皮、枣股、枣刺、枣吊、落叶、落枣等越冬组织，在室内分离镜检，确定了枣缩果病病原菌的越冬场所。在本地引起枣缩果病的病原菌是半知菌亚门中的细交链孢菌和聚生小穴壳菌，其中细交链孢菌主要越冬场所为落叶（分离率〉90％），枣刺、枣吊和落果也为重要越冬场所（分离率10％～80％），树皮、枣股上不是主要越冬场所（分离率〈10％）。     

茶薪菇多孢快速育种简报

采用担孢子多孢诱变、群体杂交与组织分离进行茶薪菇育种比较,确定简单快速的多孢育种方法。经多孢杂交,筛选出高产稳定菌株A207,该株长势快,产量高,菇形好,其鲜菇效率较原株增加16．1％,畸菇率下降3．4％。     

银耳菌分离方法的研究

用单孢分离、基本分离和组织分离获得银耳菌，银耳菌的纯培养与伴生菌长在一起成为银耳母种。在三种方法中，以单孢分离获得的银耳原基、基内菌丝与伴生菌配制的母种质量好，成功率亦高。基内分离纯培养的方法不如单孢分离的方法好，基内分离银耳菌丝和伴生菌的混合物，得到的母种质量不稳定。组织分离的方法不能采用。     

蘑菇单孢分离和诱导萌发

双孢蘑菇担子上一般仅着生两个担孢子,而不是通常的4个,因此,不易得到单核体。研究双孢蘑菇的遗传学或进行杂交育种工作,不但需将大量的孢子在无菌条件下分别萌发,而且需使一个担子上的所有孢子萌发,还要得到其减数分裂的全部产物。用通常的孢子悬液法在平板上分离很难得到单孢子分离物,因为成团的孢子会产生多孢子培养物。用显微技术进行单孢子分离费时费力,成功率也很低,分离到斜面上的单孢子萌发率一般仅1％。     

芸薹根肿菌单孢分离技术体系构建及沈阳地区根肿菌的鉴定

芸薹根肿菌严重危害大白菜和其他十字花科作物。由于根肿菌通常以多个生理小种混生状态进行侵染,导致抗病品种的抗性不能持久。以沈阳地区根肿菌为材料,进行了单孢分离技术体系构建,结果表明采取冷冻盒接菌法对两日龄大白菜幼苗进行单孢接种,可将侵染率提高到47.86%;采用基质过渡培养可将大白菜植株成活率提高到67%。利用上述根肿菌单孢分离技术体系对沈阳地区根肿菌进行单孢分离和Williams生理小种鉴定,结果表明沈阳地区根肿菌存在4号、8号、9号、11号生理小种,其中4号生理小种为优势小种。     

平菇单孢杂交育种的研究

目前,已有不少关于平菇单孢杂交育种的研究报道,其中大多是采用平板稀释涂布孢子悬液法分离单孢,并且依据锁状联合的有无来判断是否是单核菌丝,这种分离和检测方法很有可能会将多孢菌落误认为单孢菌落.平菇有性生殖为异宗配合,四极性类型.用上述方法获得的单核菌丝进行杂交时,就会出现种质不纯的严重问题.引起菌种退化,本实验通过一种简便的挑选单孢子的方法进行平菇品种间的杂交育种.     

金针菇单孢结实性的研究及其应用

本研究采用稀释镜检法对金针菇的６个品种，共计１８９８个担孢子分离培养，结果检测出１４４个单孢结实菌株。这些菌株与其它非单孢结实菌株支配，筛选出６个强优组合。     

Protective effect of aFGF delivery by ultrasound-targeted microbubble destruction on left ventricular function in diabetic cardiomyopathy rats

AIM：To observe the protective effect of delivery of acidic fibroblast growth factor (aFGF) to myocardium by ultrasound-targeted microbubble destruction (UTMD) on left ventricular function in diabetic cardiomyopathy (DCM) rats and to investigate the possible mechanisms. METHODS：Twenty-four rats were intraperitoneally injected with streptozocin to induce DCM and were randomly divided into DCM group and aFGF treatment group. Twelve healthy rats served as normal controls. The rats in aFGF treatment group were infused with SonoVue-aFGF mixed fluid through tail vein and UTMD was simultaneously performed. Four weeks after intervention, all rats underwent cardiac catheterization to mea-sure left ventricular end-systolic pressure (LVESP), left ventricular end-diastolic pressure (LVEDP) and the maximal increase/decrease rate of left ventricular pressure (LV±dp/dtmax). The microvessel density (MVD) of rat myocardial tissues was measured by immunohistochemical staining for CD31. The myocardial collagen volume fraction (CVF) was determined by improved Masson staining. The apoptotic index (AI) was detected by TUNEL method. RESULTS：Four weeks after intervention, the LVESP and LV±dp/dtmax in aFGF treatment group were significantly increased compared with DCM group (P<0.01), while the LVEDP in aFGF treatment group was significantly lower than that in DCM group (P<0.01). The MVD in aFGF treatment group was significantly increased compared with DCM group (P<0.01), but the CVF and AI in aFGF treatment group were significantly lower than those in DCM group (P<0.01). CONCLUSION: Delivery of aFGF to diabetic myocardium by UTMD could improve the left ventricular function of DCM rats and may be a new feasible therapeutic method for DCM.     

Effects of pentoxifylline on ventricular remodeling and cardiac function of dilated cardiomyopathy rats

AIM: To explore the effects of pentoxifylline (PTX) on ventricular remodeling and cardiac function in dilated cardiomyopathy (DCM) rats.METHODS: Lewis rats were randomly allocated to a myocin-induced dilated cardiomyopathy (DCM) group receiving saline (n=10), a DCM group receiving PTX (PTX group; 25 mg·kg^-1·d^-1, ip, for 30 days, n=10) or healthy control group (n=10). The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-10 in the blood plasma were analyzed by ELISA. The extent of fibrosis was estimated using Massons staining and immunohistochemistry analyses. Cardiac structure and function were measured by echocardiography.RESULTS: PTX decreased plasma levels of TNF-α and IL-6, and increased IL-10 level in DCM animals compared with DCM group ［TNF-α: (7.21±0.24) μg/L vs (19.30±1.31) μg/L, P<0.01; IL-6: (119.60±36.58) ng/L vs (189.50±13.25) ng/L, P<0.05; IL-10: (41.26±3.27) μg/L vs (32.45±4.32) μg/L, P<0.05］. Collagen volume fraction (CVF), perivascular collagen area (PVCA) and collagen Ⅰ/Ⅲ ratio were lower in PTX group than those in DCM group ［CVF: (16.45±3.01)% vs (23.33±4.43)%, P<0.05; PVCA: 4.58±2.10 vs 13.74±4.29, P<0.05; Ⅰ/Ⅲ ratio: 2.84±0.67 vs 4.22±0.54, P<0.01］. Left ventricular end-diastolic dimension reduced ［(6.11±0.51) mm vs (6.46±0.28) mm, P<0.05］ and left ventricular ejection fraction elevated ［(77.29±5.20)% vs (62.73±10.11)%, P<0.01］ by PTX compared with DCM.CONCLUSION: PTX modulates plasma levels of inflammatory cytokines, delays the ventricle remodeling and improves the heart function in DCM rats.     

Relationship between 3-nitrotyrosine and myocardial cell apoptosis in rat diabetic cardiomyopathy

AIM: To explore the relationship between 3-nitrotyrosine (3-NT) level in hearts or blood and myocardial cell apoptosis in rat diabetic cardiomyopathy (DCM). METHODS: Sixty Sprague-Dawley (SD) rats (male, 8-week-old) were randomly divided into 4 groups: normal group, diabetic cardiomyopathy group (DCM group), diabetic rats treated with valsartan (40 mg·kg^-1·d^-1, D+V group) and DCM rats treated with valsartan (40 mg·kg^-1·d^-1, DCM+V group). Apoptotic index (AI) of rat cardiac myocytes was examined by TUNEL. The expression index (EI) of 3-NT in rat cardiac myocytes was examined by immunohistochemistry. The 3-NT concentration in rat serum was examined by ELISA. RESULTS: (1) Significant differences of the heart weight indexes among the 4 groups were observed (P<0.01). The heart weight indexes in DCM group and DCM+V group were higher than those in normal group and D+V group (P<0.01). (2) The EI of 3-NT in the cardiac myocytes was positively correlated with the AI of the cardiac myocytes in the same group (P<0.01), but the concentration of 3-NT in blood had no correlation with the AI of cardiac myocytes (P>0.05). (3) The difference of AI of cardiac myocytes among the 4 groups had statistical significance (P<0.01). The arrangement from high to low of AI was DCM group > D+V group and DCM+V group > N group (P<0.05). (4) The EI of 3-NT in DCM group was the highest as compared to other groups (P<0.05). (5) No statistical difference of 3-NT concentration in blood among the 4 groups was observed (P>0.05). CONCLUSION: (1) The expression of 3-NT in DCM myocardial tissues in SD rats is significantly increased and closely correlated with the apoptosis in myocardial cells. Valsartan inhibits 3-NT expression in DCM myocardial cells, thus inhibits the DCM myocardium apoptosis. (2) The 3-NT level in blood can not be true for reflection of 3-NT expression in DCM myocardial tissues and its effect on myocardial cell apoptosis.     

Synergistic effect of insulin and selenium in combination on inhibiting myocardial apoptosis in rats with diabetic cardiomyopathy

AIM: To investigate the effect of insulin and selenium in combination on the apoptosis and the expression of Ku70, acetylated Ku70, Bax and cytochrome C in myocardial cells of the rats with diabetic cardiomyopathy(DCM), and to explore the mechanism of insulin and selenium in their synergistic anti-DCM effect. METHODS: SD rats (n=50) were randomly grouped into control, DCM, DCM with insulin treatment (DCM+In) group, DCM with selenium treatment (DCM+Se) group, and DCM with insulin and selenium combination treatment (DCM+In+Se) group. Mitochondrial membrane potential (MMP) was measured by flow cytometry. The cell apoptosis was observed by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). The levels of Ku70, Bax and cytochrome C were examined by Western blot. The acetylation status of Ku70 was detected by co-immunoprecipitation.RESULTS: The rats in DCM group showed marked cell apoptosis compared with the control rats. The levels of Ku70 and acetylated Ku70 declined significantly compared with control group. Bax significantly translocated from cytoplasm into mitochondria and cytochrome C translocated from mitochondria into cytoplasm compared with control group. Compared with DCM+In group or DCM+Se group, insulin and selenium in combination significantly inhibited the apoptosis, down-regulated Ku70 and acetylated Ku70 levels, and prevented Bax and cytochrome C translocation.CONCLUSION: Insulin and selenium synergistically inhibits myocardial apoptosis by regulating Ku70 acetylation and inhibiting Bax translocation.     

柑橘落花落果的营养元素含量及其脱落损耗

 以成年纽荷尔脐橙、兴津温州蜜柑和沙田柚为材料,研究落蕾、落花和脱落幼果中的养分 含量及其脱落造成的损耗。结果表明,3 个品种的蕾和花营养元素含量为N 3.34% ~ 3.66%,P 0.26% ~ 0.31%,K 1.67% ~ 2.30%,Ca、Mg 和S 含量0.17% ~ 0.48%,Fe、Mn、Zn、Cu 和B 含量8.1 ~ 87.5 mg · kg^-1。 除花的B 含量显著高于蕾外,花和蕾的多数营养元素含量相近,且品种间差异小。与蕾和花相比,幼果 的N 和P 含量较少,N 和P 含量分别少21.8%和19.5%,但Fe 含量较多,其它营养元素含量变化不明显。 纽荷尔脐橙、兴津温州蜜柑和沙田柚单株脱落的蕾、花和幼果干物质量合计分别为3 016.3 g、3 533.6 g 和1 486.7 g,由此所造成的单株主要养分损耗为N 49.2 ~ 119.4 g、P 4.3 ~ 10.1 g、K 30.1 ~ 76.2 g、Mg 2.5 ~ 7.1 g、Zn 24.0 ~ 81.5 mg 和B 65.5 ~ 170.2 mg。兴津温州蜜柑和沙田柚均以落花的养分损耗最大,落蕾其 次,落果最小;纽荷尔脐橙则是落花和落蕾的养分损耗相近且远高于落果。品种间以兴津温州蜜柑脱落 的养分损耗最大,纽荷尔脐橙其次,沙田柚最小。因此,柑橘应控制过量开花,并在萌芽开花期及时补 充养分。     

Regulatory effect of miR-146 on diabetic cardiomyopathy

AIM: To investigate the role of microRNA-146 (miR-146) in the pathogenesis of diabetic cardiomyopathy (DCM), and to observe the expression levels of miR-146 and its downstream target genes. METHODS: Male C57BL/6 mice (n=60) were randomly divided into experimental group (DCM, n=30) and control group (control, n=30). The mice in DCM group were intraperitoneally injected with low dose (50 mg/kg) of streptozo-tocin (STZ) to induce diabetic myocardial model, and the mice in control group were given intraperitoneal injection of citrate buffer. At the end of 12 weeks, the hearts were removed, HE and Masson staining were performed to observe the cardiac pathological changes. RT-qPCR were used to detect the expression of miR-146 a and miR-146b and the the mRNA expression of related downstream genes interleukin-1 receptor-associated kinase 1 (IRAK1) and TNF receptor-associated factor 6 (TRAF6). Western blot was used to determined the protein levels of IRAK1 and TRAF6. RESULTS: At the end of 12 weeks, HE staining showed the hypertrophy and structural disorder of the myocardial cells in DCM group. Masson staining showed that the collagen fibers of myocardium were increased in DCM group; RT-qPCR showed that the levels of miR-146a and miR-146b in DCM group were significantly decreased compared with control group (P<0.01). The mRNA levels of IRAK1 was increased (P<0.01), and the mRNA levels of TRAF6 was declined (P<0.01). Western blot showed that compared with control group, the protein expression of IRAK1 was increased in DCM group, and the protein expression of TRAF6 was decreased with statistically significant.CONCLUSION: miR-146 may involve in the occurrence and development of diabetic cardiomyopathy by regulating inflammatory reactions and targeting IRAK1.     

Cytokine production in mice with experimental cardiomyopathy treated with anti-L3T4 monoclonal antibody at different stages

AIM: To clarify the mechanism of treating autoimmune cardiomyopathy at different stages with anti-L3T4 monoclonal antibody. METHODS: Mice immunized with human mitochondria ADP/ATP peptides were used as the cardiomyopathy (DCM) group, and the sham-immunized mice were regarded as the controls. Mice receiving early treatment were immunized with the same peptides, followed by the injection of 400 μg of anti-L3T4 on day 0, 1 and 2 post-immunization. Mice in the late treatment group were immunized as of the early treatment group but anti-L3T4 was administered 3 months post-immunization. The cytokine expression was measured with three-color flow cytometry to quantitate the splenic Th1/Th2 cell subsets in the different groups of mice. In addition, serum and myocardial cytokines were measured by enzyme-linked immunosorbent assay and real-time PCR. RESULTS: Th1 and Th2 subsets in the early treatment group were similar to those in control group, but were drastically lower than those in DCM group. Mice in the late treatment group showed an increased level of Th1-related cytokines, while the Th2 level was between the DCM and early treatment group. IFN-γ and IL-6 levels in early treatment group were similar to those in control group. In the early treatment group, IL-4 level was higher than that in control and lower than that in DCM group, whereas IL-2 and TNF-α contents were lower than those in control and DCM group. In the late treatment group, IFN-γ and IL-2 levels were higher than those in DCM group and lower than those in the early treatment group, while IL-6 and IL-4 levels were lower than those in DCM group. CONCLUSION: These results suggest that the cytokine production in cardiomyopathic mice may be repressed by treatment with anti-L3T4 at different stages. Early treatment with anti-L3T4 has better inhibitory function than treatment in late stage of autoimmune cardiomyopathy.     

Protective effect of DIZE on heart function of rats with diabetic cardiomyopathy

AIM: To observed the protective effect of diminazene aceturate (DIZE), an angiotensin-converting enzyme 2 (ACE2) activator, on rats with diabetic cardiomyopathy (DCM). METHODS: Male Wistar rats (n=30) were randomly divided into normal control group, DCM group and DIZE treatment group (DIZE group). The rats in DCM group and DIZE group were intraperitoneally injected with streptozotocin (65 mg/kg) to establish diabetic model. After 12 weeks, the diabetic rats were infused with DIZE at 15 mg·kg^-1·d^-1 or the same volume of saline for 4 weeks using osmotic minipump. The cardiac function was measured at the end of the 16th week. The methods of Mason staining and HE staining were used to observe the morphological changes of the myocardial tissue. Western blot, ELISA and immunohistochemistry were used to observe the changes of ACE2, angiotensin (Ang)Ⅱ, Ang-(1-7), interleukin (IL)-1, IL-6 and connective tissue growth factor (CTGF). RESULTS: DIZE significantly improved the expression of ACE2 in diabetic rats (P<0.05). Compared with DCM group, the levels of IL-1 and IL-6 in DIZE group were significantly decreased, and the cardiac function in DIZE group was significantly improved (P<0.05). CONCLUSION: ACE2 endogenous agonist DIZE significantly increases the ACE2 level and reduces the level of inflammation, thus protecting the heart function of DCM rats.     

Abnormal T-cell receptor signal pathway in murine autoimmue cardiom yophy induced with adenine nucleotide translocase

AIM: To explore the molecular mechanism in the pathogenesis of dilated cardiomyopathy (DCM) by analyzing the expression of T cell signaling molecules in mice with autoimmune DCM. METHODS: Mouse DCM model was induced by immunizing the animals with adenine nucleotide translocase (ANT) synthetic peptides. P56lck in T cells was detected with real-time fluorescent quantitative PCR in both DCM-group and the sham-immunized controls. At the same time, flow cytometry was used for quantity of Th cell intracellular cytokine IFN-γ and IL-4, ELISA for examining the level of serum anti-ANT antibody, immune histochemistry for investigating the expression of CD45 in Th cells. RESULTS: The mRNA expression of P56lck (1 369.51±874.05 vs 47.93±10.21, P<0.01), the percentage of IFN-γ and IL-4 (especially IL-4) (8.27±1.29 vs 5.58±0.59, P<0.01; 9.93±1.53 vs 2.05±0.21, P<0.01), the level of anti-ANT autoantibody (0.105±0.015 vs 0.006±0.002, P<0.01) and expression of CD45 (0.154±0.021 vs 0.026±0.008, P<0.01) were all elevated significantly in DCM-group compared with the controls. CONCLUSION: Impairment of T cell receptor (TCR) signal transduction pathway plays an important role in the development of DCM induced with ANT synthetic peptides in mice.