暑期教学实习记事——《草人诗记》之二

高山望月小,夜风来远山。师生同薄帐,互道不觉寒。瓶水恐冻裂,拥被相与眠①。晨起忙炊事,浓烟煮面团。切菜用剪刀②,调味只有盐。捧碗暖冻手,热气凝眼睑③。天冷心中热,学子恋草原。

《草人诗记》开场白

开宗明义，先说明来历。两年以前，我中断了连续五年的《草业琐谈》专业杂文。热心的同道们建议赓续。我也想与读者恢复交流通道。但年事日高，仍循《草业琐谈》旧轨，日常工作之余，每月一篇短文，怕精力有所未逮，于是想到利用旧有诗作，配以简略文字，以记述某些新旧故事，也不失为一种文化记忆。这就是《草人诗记》。

饥寒无惧伴熊狼——《草人诗记》之一

月夜清辉漫山梁，溪畔孤帐泛青光。夜闻狼嚎传莽野，晨看熊掌绕帐房。浓烟滚滚难为炊，寒风瑟瑟透衣裳。薄帐一顶雪地居，饥寒无惧伴熊狼。

1978年科学大会有感三首——《草人诗记》之七

之一 草虫僵卧闻惊雷，且舒节肢试地温。 一种生意生革翅，原来大地已回春。

草人足迹——《草人诗记》之十一

草人挟囊走荒谷，带泥足迹没丛芜①。 最是寂寞含悲日，草地农业有若无②。

草人形状——《草人诗记》之八

2009年12月27日，深圳到北京的飞机上，我翻阅唐诗。王昌龄有《塞上曲》云：“蝉鸣空桑林，八月萧关道。出塞复入塞，处处黄芦草。从来幽并客，皆向沙场老。莫学游侠儿，矜夸紫骝好”。读诗至此，我掩卷叹日，这不正是含蕴了草人形状吗？遂步原韵和诗一首。

Induction of apoptosis and G2/M cell cycle arrest by oridonin in human gastric cancer BGC-823 cells

Aim To investigate in vitro apoptosis-induction effects of oridonin on gastric tumor cells BGC-823 and its effects on cell cycle, mitochondrial membrane potential and intracellular Ca^2＋ to shed light on the mode of its anticancer action. Methods The MTT method was used to investigate the inhibitory effect of oridonin on BGC-823 cells. The apoptosis-induction effect was evaluated by confocal laser microscopy and flow cytometry. The change of mitochondrial membrane potential and the increase of intracellular Ca^2＋ were assessed by fluorescence probe rhodamine123 and Fluo 3-AM, respectively, with flow cytometry. The expression of apoptosis and cell cycle related proteins was studied using western blotting. Results Oridonin inhibited BGC-823 cells growth with IC50 of 22.21 p, mol.L^-1. It induced apoptosis in a dose-dependent manner. In addition, it decreased mitochondria membrane potential, increased intracellular Ca^2＋, and activated pro-caspase 3. BGC-823 cells were arrested in G2/M cell cycle phase with lower expression of cyclin A protein. The up-regulation of p53 was observed before apoptosis and cell cycle arrest occurred. Conclusion Oridonin inhibits the proliferation of BGC-823 cells through G2/M cell cycle arrest and apoptosis induction, which is mediated by influx of Ca^2＋, up-regulation of p53, activation of caspase-3, and down-regulation of cyclin A.

Effects of Shikonin on Proliferation, Apoptosis, and Extracellular Matrix of Human Mesangial Cells

Objective:To investigate the effect of shikonin on the proliferation, apoptosis and extracellular matrix (ECM) of human mesangial cells (MC). Methods: MC was cultured in vitro with different concentrations of glucose (30, 50, 80 mmol/L). The cell growth was observed by using MTT method and apoptosis by using an aunexin-V-Fluos. Immunohistochemical studies for Laminin (LN), Fibronectin (FN) and type Ⅳ Collagens (Col Ⅳ) were measured. Results: Shikonin inhibited their growth (P<0.05) and apoptosis in the glycated cultured cells. Shikonin 0.05 mmol/L significantly reduced the secretion of LN, FN and Col Ⅳ from MC (P<0.05) cultured in 30, 50 and 80 mmol/L glucose. Conclusion: Shikonin could prevent or treat diabetic nephropathy (DN) and glomerulosclerosis (GS).

The Nucleosides Contents and Their Variation in Natural Cordyceps sinensis and Cultured Cordyceps Mycelia

Aim: To compare the contents of nucleosides from natural Cordyceps sinensis and cultured Cordyceps mycelia, and to study the effect of humidity and heat on the content of nucleosides. Methods: The contents of nucleosides were determined by using high performance capillary electrophoresis (HPCE). Beckman P/ACE System 5010 apparatus equipped with a UV detector and a Beckman untreated fused-silica capillary (57 cm 75 mm, 50 cm effective length) was used. Before sample injection, the capillary was rinsed with 1 molL-1 sodium hydroxide solution and running buffer for 5 min, respectively. A voltage of 20 kV was applied for the separation. Pressure injection was 586 kPa for 6 seconds, and the wavelength of detector was 254 nm. The running time was 20 min at 20 oC. The effect of humidity and heat on the contents of nucleosides from natural Cordyceps sinensis and cultured Cordyceps mycelia was observed for 1, 3, 5 and 10 days at temperature 40 oC, and relative humidity 75%. Results: The content of nucleosides from natural Cordyceps sinensis was higher than that from cultured Cordyceps mycelia. But the contents of nucleosides from freshly collected natural Cordyceps sinensis were very low, even below the limit of quantitation. The contents of nucleosides from natural Cordyceps sinensis were significantly increased by humidity and heat, but this phenomenon was not observed in cultured Cordyceps mycelia. Conclusion: There are differences between the nucleosides from natural Cordyceps sinensis and cultured Cordyceps mycelia. The nucleosides in natural Cordyceps sinensis may be derived from the degradation of nucleic acids. This implies that adenosine being used for the quality control of natural Cordyceps sinensis may have to be reconsidered.

Changes of Monoanine Levels in Different Brain Regionsof Rat with Ventral GIobus Pailidus Injured by KainicAcid Injection

Ventral globus pallidus-injured rats by kainic acid(10 mg) were used to derter-mine the monoamine levels in four different brain regions.In this model,a great decrease of NE con-centration was observed in both hippocampus and frontal cortex compared with nonnal contro.This result hints that a damage of noradrenergic neurons in this model has occurred. DA concentrations in the four brain zones after kainic acid injection were all reduced, but only in the frontal cortex and striatum the pronounced reductions were discovered while DA turnover rates in frontal cortex, stria-tum and meddullapons were significantly reduced. These results revealed a DA metabolic disorder occurring in these regions.However,5-HT concentrations as well as DBH activity, expressed by ratio of NE/DA,showed no marked difference in this model. In our study it is found that the changes of monoamine levels in this model basically reflect those discovered in AD patients.

Development of Artificial Diet for Erigonidium graminicolum(sundevall)

[Objective] This study was to find out the artificial diet formula for Erigoni- dium graminicolum （Sundevall） to realize its indoor artificial breeding. [Method] Using orthogonal decomposition method, the diet most suitable for the normal and quick growth of Erigonidium graminicolum （sundevall）, was developed by adding different species and different proportions of fresh insects to the artificial diet based on the formula of total nutrient solution proposed by Zhao Jing-zhao. [Result] The most suit- able formula for the development and growth of Erigonidium graminicolum （Sundevall） is 10 ml of egg white, 2 g of honey, 2 g of sucrose, 10 ml of water, 5 g of Ci- cadella viridis, moderate amount of cornmeal, compound vitamin B1 and vitamin C1. [Conclusion] The development of new type artificial diet can provide timely and nec- essary material premise for its large-scale indoor propagation.

Study on the Degradation Trend of Ryegrass-White Clover Artificial Grassland

[Objective] The study aimed to explore the degradation law and trend of artificial grassland. [Method] Taking the ryegrass （Lolium perenne） - white clover （ Trifolium repens） artificial grassland in Maiping Township, Guizhou Province as the research object, the grassland vegetation of 40 quadrate from different areas （area around the sheep shed, hilltop, hillside, flatland at the foot of the hill） were analyzed by comparing the dominance and richness index. [ Result] Degradation of different degrees appeared in various areas of this artificial grassland. To be specific, the degradation of grassland around the sheep shed was not serious for less consumption by the sheep; however, the areas on the hilltop and hillside degenerated to the grassland types of dallis grass （ Paspalum dilatatum Poir. ） -Indian lovegrass （ Eragrostis pilosa） and dallis grass - cogongrass [ Imperata cylindrica （ Linn. ） Beauv. ], respectively, and the area at the foot of the hill degenerated to the grassland type dominated by garland chrysanthetnum （ Chrysanthemum coronarium L. ） and knotgrass （ Paspalum distichum L. ）. [ Conclusion ] This study provided a basis for grassland improvement as well as the efficient and sustainable utilization of grazing-type artificial grassland in South China.

Patent Information Analysis of Front-end Industry Chain in China's Tobacco Industry

The research performed analysis on patent applicant ranking, IPC, patent type and legal status in order to explore application and authorization of front-end industry chain dominated by planting, breeding, soil improvement and substrate culti- vation technologies. The results showed that research institutions and big tobacco enterprises play a leading role in the tobacco industry, for example, Yunnan Tobac- co Agricultural S＆T Research Institute and Guizhou Tobacco Agricultural S＆T Re- search Institute have a total of 26 patents; A01G subclass represents 58% of total patents, dominated by Yunnan Tobacco Agricultural S＆T Research Institute and Henan Agricultural University; there are only invention and practical use patents, of which invention patents represent 92%; authorized patents take up to 31% and in- valid patents represent 23%.

Soil CH_4 fluxes response to understory removal and N-fixing species addition in four forest plantations in Southern China

CH4 is one of the most important greenhouse gases, and mainly comes from soils in forest ecosystems. The objective of this study was to determine the effects of forest management practices such as understory removal and N-fixing species （Cassia alata） addition, on soil CH4 fluxes in four forest plantations in southern China. Fluxes of CH4 were measured in Eucalyptus urophylla plantation （B1）, Acacia crassi-carpa plantation （B2）, 10-native-species-mixed plantation （B3）, and 30-native-species-mixed plantation （B4） stands using the static chamber method in Southern China. Four forest management treatments, includ-ing （1） understory removal and replacement with C. alata （UR＋CA）; （2） understory removal only （UR）; （3） C. alata addition only （CA）; and （4） control without any disturbances （CK）, were applied in the four forest plantations. The results showed that plantation types had a significant effect on soil CH4 fluxes. B1 and B2 tended to be CH4 consumers, while B3 and B4 inclined to be CH4 producers. UR decreased CH4 fluxes by providing a more optimal soil temperature and moisture regime for mi-croorganism community and increasing substrate mineralization. How-ever, CA enhanced CH4 fluxes in B1 and B2 for N-fixing function of C. alata while lowered CH4 fluxes in B3 and B4. Soil CH4 flux rate was significantly related to soil temperature and moisture conditions in the top 10-cm soil layer. Furthermore, the quality of substrates, such as Soil Organic Carbon （SOC） and mineral N might also be important driving factors for CH4 fluxes. This study improved our understanding on CH4 fluxes in plantations under different management practices such as UR and CA.

Preliminary Studies on High-performance Liquid Chro-matography Chemiluminescence Determination of theSaturated Fatty Acids(C_(16) C_(18))in Human Serum

The peroxyoxalate chemiluminescence(CL)detection of fatty acids in human se- rum combined with high-performance liquid chromatography (HPLC)is described.Some fatty acids in serum were extracted with a 1 :1(v/v)mixture of chloroform-n-heptane.2-(4-Hydrazinocarbonyl- phenyl)-4,5-diphenylimidazole (HCPI) was used as a fluorescent labelling reagent of the fatty acids. The labelling reaction was carried out at 30℃ for 1 h at pH 6.5 and the resulting reaction mixture was sudjected to HPLC. The labelled fatty acid C_(17)(P-C_(17))was used as the internal standard. The la- belled fatty acids C_(16) and C_(18) were separated within 18 min on an ODS-8OTM column (150 mm× 6 mm ID,5μm,Tosoh Japan).The calibrlation curves of fatty acids from the spiked control serum were Y_1=-0.003 7 + 0.0028X_1,r=0.994 for FA C_( 16) and Y_2=0.00 1 2 + 0.00098X_2,r=0.999 for FA C_( 18),respectively.The average recoveries of facids from the spiked contrl serum were 107.2%(n=8,RSD=4.3%)for FA C_(16) and 97.35%(n=8, RSD=4.0%)for FA C_(18),respectively.The lower detection limits of fatty acids after reaction were 12μmol per 20μl injection for FA C_(16) and 18 μmol per 20μl injection for FA C_(18),respectively(signal to noise ratio, S/N=2).The HPLC/CL method was applied to the determination of FA C_(16) and FA C_(18) in normal human serum and the results showed that the concentrations of fatty acids in normal human serum were 0.134 ± 0.009 μ mol/ml serum(n=5) for FA C_(16) and 0.052±0.028 μmol/ml serum(n=5)for FA C_(18),respectively.

Present Situation in Preventing and Treating Liver Fibrosis with TCM Drugs

Considerable evidences have shown that the mechanism of TCM drugs for preventing and treating liver fibrosis is very complicated. TCM treatment can not only inhibit viral replication, ameliorate inflammation and promote blood circulation in the liver, and enhance regeneration of the hepatic cells, but also inhibit HSC proliferation, intra- and extracellular secretion, decrease the secretion of collagen and promote its degradation and re-absorption. However, most of the animal models are only suitable for studies of acute hepatitis. Establishment of cell lines suitable for studies of fibrosis is still at its initial stage. What we expect is that comprehensive clinical studies in TCM treatment of liver fibrosis will be carried out and evaluation of each datum given, both of which are of importance.

Advances in TCM Treatment of Primary Hepatocarcinoma

Characterized by insidious onset,fast progress,wide epidemic with poor therapeutic effect and shortsurvival period,primary hepatocarcinoma is one ofthe malignant tumors that threatens the human health.Rich experiences and gratifying achievements havebeen obtained by Chinese medical workers.

Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Clinical Observation on Treatment of NonParvicellular Carcinoma of the Lung with Jin Fu Kang Oral Liquid

Jin Fu Kang Oral Liquid ([symbol: see text]), made of traditional Chinese drugs for supplementing qi and nourishing yin, was developed according to the common symptoms in lung carcinoma with deficiency of both qi and yin. Of the 96 cases in the Jin Fu Kang group, 1 case got complete remission (CR) after treatment, 8 cases partial remission (PR), 52 cases no change (NC), PR + NC covering 63.5%. Of the 52 cases in the group of Jin Fu Kang plus chemotherapy, 11 cases got PR after treatment, 26 cases NC, PR + NC covering 71.2%. Of the 25 cases in the chemotherapy group, 4 cases got PR after treatment, 11 cases NC, PR + NC covering 60.0%. The results show that the therapeutic effectiveness in the Jin Fu Kang group and the group of Jin Fu Kang plus chemotherapy was better than that in the chemotherapy group. The one-year survival rate and the two-year survival rate after treatment in the Jin Fu Kang group were 67.3% and 67.3% respectively; 66.7% and 66.7% in the group of Jin Fu Kang plus chemotherapy; and 40.3% and 0.0% in the chemotherapy group. The improvement of clinical symptoms, increase of body weight and improvement of health situation (KPS marks) after treatment in both the Jin Fu Kang group and the group of Jin Fu Kang plus chemotherapy were better than that in the chemotherapy group. Some indicators of immunology and hemogram after treatment were greatly improved in the Jin Fu Kang group, worse in the chemotherapy group, but no obvious improvement in the group of Jin Fu Kang plus chemotherapy.

The Inhibitory Effect of Oridonin on the Growth of Fifteen Human Cancer Cell Lines

OBJECTIVE To study the inhibitory effect of oridonin on the growth of cancer cells. METHODS Fifteen human cancer cell lines were subjected to various concentrations of oridonin in culture medium. The inhibitory rate of cell growth was measured by the MTT assay, and compared with a negative control and 5-Fu-positive control. RESULTS The 50% inhibiting concentration （IC50） and maximal inhibition （Imax） of oridonin shown by studying the growth of the cancer cell lines were as follows： leukemias （HL60 cells： 3.9 μg/ml and 73.8%, K562 cells： 4.3 μg/ml and 76.2%）; esophageal cancers（SHEEC cells： 15.4 μg/ml and 99.2%, Eca109 cells： 15.1 μg/ml and 84.6%, TEl cells： 4.0 μg/ml and 70.2%）; gastric cancers （BGC823 cells： 7.6 μg/ml and 98.7%, SGC7901 cells： 12.3 μg/ml and 85.7%）; colon cancers （HT29 cells： 13.6 μg/ml and 97.2%, HCT cells： 14.5 μg/ml and 96.5%）; liver cancers （Bel7402 cells： 15.2 μg/ml and 89.2%, HepG2 cells： 7.1 μg/ml and 88.3%）; pancreatic cancer （PC3 cells： 11.3 μg/ml and 68.4%）; lung cancer （A549 cells： 18.6 μg/ml and 98.0% ）; breast cancer （MCF7 cells： 18.4 μg/ml and 84.7%）; uterine cervix cancer （Hela cells： 13.7 μg/ml and 98.5%）. CONCLUSION Oridonin had a relatively wide anti-tumor spectrum, and a relatively strong inhibitory effect on the growth of the 15 human cancer cells. Inhibitory effects were concentration dependent.