[Huazhi Rougan Granules attenuates steatosis in cell model of nonalcoholic fatty liver disease by inducing autophagy].

To investigate the effect of Huazhi Rougan Granules(HZRG) on autophagy in a steatotic hepatocyte model of free fatty acid(FFA)-induced nonalcoholic fatty liver disease(NAFLD) and explore the possible mechanism. FFA solution prepared by mixing palmitic acid(PA) and oleic acid(OA) at the ratio of 1∶2 was used to induce hepatic steatosis in L02 cells after 24 h treatment, and an in vitro NAFLD cell model was established. After termination of incubation, cell counting kit-8(CCK-8) assay was performed to detect the cell viability; Oil red O staining was employed to detect the intracellular lipid accumulation; enzyme-linked immunosorbnent assay(ELISA) was performed to measure the level of triglyceride(TG); to monitor autophagy in L02 cells, transmission electron microscopy(TEM) was used to observe the autophagosomes; LysoBrite Red was used to detect the pH change in lysosome; transfection with mRFP-GFP-LC3 adenovirus was conducted to observe the autophagic flux; Western blot was performed to determine the expression of autophagy marker LC3B-Ⅰ/LC3B-Ⅱ, autophagy substrate p62 and silent information regulator 1(SIRT1)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK) signaling pathway. NAFLD cell model was successfully induced by FFA at 0.2 mmol·L~(-1) PA and 0.4 mmol·L~(-1) OA. HZRG reduced the TG level(P<0.05, P<0.01) and the lipid accumulation of FFA-induced L02 cells, while elevated the number of autophagosomes and autophagolysosomes to generate autophagic flux. It also affected the functions of lysosomes by regulating their pH. Additionally, HZRG up-regulated the expression of LC3B-Ⅱ/LC3B-Ⅰ, SIRT1, p-AMPK and phospho-protein kinase A(p-PKA)(P<0.05, P<0.01), while down-regulated the expression of p62(P<0.01). Furthermore, 3-methyladenine(3-MA) or chloroquine(CQ) treatment obviously inhibited the above effects of HZRG. HZRG prevented FFA-induced steatosis in L02 cells, and its mechanism might be related to promoting autophagy and regulating SIRT1/AMPK signaling pathway.

[Mechanism of Huangjing Qianshi Decoction in treatment of prediabetic mice based on transcriptome sequencing].

Based on transcriptome sequencing technology, the mouse model of prediabetes treated with Huangjing Qianshi Decoction was sequenced to explore the possible mechanism of treating prediabetes. First of all, transcriptome sequencing was performed on the normal BKS-DB mouse group, the prediabetic model group, and the Huangjing Qianshi Decoction treatment group(treatment group) to obtain differentially expressed genes in the skeletal muscle samples of mice. The serum biochemical indexes were detected in each group to screen out the core genes of Huangjing Qianshi Decoction in prediabetes. Gene Ontology(GO) database and Kyoto Encyclopedia of Genes and Genomes(KEGG) database were used to conduct signaling pathway enrichment analysis of differentially expressed genes, and real-time quantitative polymerase chain reaction(RT-qPCR) was used to verify them. The results showed that the levels of fasting blood glucose(FBG), fasting insulin(FINS), insulin resistance index(HOMA-IR), total cholesterol(TC), triglycerides(TG), and low-density lipoprotein cholesterol(LDL-C) in the mouse model were significantly decreased after treatment with Huangjing Qianshi Decoction. In the results of differential gene screening, there were 1 666 differentially expressed genes in the model group as compared with the normal group, and there were 971 differentially expressed genes in the treatment group as compared with the model group. Among them, interleukin-6(IL-6) and NR3C2 genes, which were closely related to the regulation of insulin resis-tance function, were significantly up-regulated between the model group and the normal group, and vascular endothelial growth factor A(VEGFA) genes were significantly down-regulated between the model group and the normal group. However, the expression results of IL-6, NR3C2, and VEGFA genes were adverse between the treatment group and the model group. GO functional enrichment analysis found that the biological process annotation mainly focused on cell synthesis, cycle, and metabolism; cell component annotation mainly focused on organelles and internal components; and molecular function annotation mainly focused on binding molecular functions. KEGG pathway enrichment analysis found that it involved the protein tyrosine kinase 6(PTK6) pathway, CD28-dependent phosphoinositide 3-kinase/protein kinase B(PI3K/AKT) pathway, p53 pathway, etc. Therefore, Huangjing Qianshi Decoction can improve the state of prediabetes, and the mechanism may be related to cell cycle and apoptosis, PI3K/AKT pathway, p53 pathway, and other biological pathways regulated by IL-6, NR3C2, and VEGFA.

[Mechanism of Huangjing Qianshi Decoction in treatment of prediabetes based on network pharmacology and molecular docking].

This study analyzed the molecular mechanism of Huangjing Qianshi Decoction(HQD) in the treatment of prediabetes based on network pharmacology and molecular docking. The active components of HQD were identified and screened based on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP, http://Lsp.nwu.edu.cn/tcmsp.php) and then the targets of the components and the genes related to prediabetes were retrieved, followed by identifying the common targets of the decoction and the disease. The medicinal component-target network was constructed by Cytoscape to screen key components. The protein-protein interaction(PPI) network was established by STRING and hub genes were identified by Cytoscape-CytoNCA, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) of the hub genes with R-clusterProfi-ler. Thereby, the possible signaling pathways were predicted and the molecular mechanism was deduced. A total of 79 active components of HQD and 785 diabetes-related targets of the components were screened out. The hub genes mainly involved the GO terms of tricarboxylic acid cycle, peptide binding, amide binding, hydrolase activity, and kinase activity regulation, and the KEGG pathways of AGE-RAGE signaling pathway, TNF signaling pathway, AMPK signaling pathway, IL-17 signaling pathway, and insulin signaling pathway. Western blot result showed that HQD-containing serum significantly reduced the expression of AKT1, AGE, and RAGE proteins in insulin resistance model cells. HQD's treatment of prediabetes is characterized by multiple pathways, multiple targets, and multiple levels. The main mechanism is that the components zhonghualiaoine, baicalein, kaempferol, and luteolin act on AKT1 and inhibit the AGE-RAGE axis.

[Pharmacodynamic mechanism of Tibetan medicine Liurui Capsules on experimental autoimmune uveitis in rats based on network pharmacology and animal experiments].

By integrating network pharmacology and animal experiments, we studied the pharmacodynamic mechanism of the Tibetan medicine Liurui Capsules in the treatment of experimental autoimmune uveitis(EAU). The active ingredients and targets of Liurui Capsules were searched against the Encyclopedia of Traditional Chinese Medicine(ETCM), Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM), and relevant literatures. The EAU-related targets were obtained from Gene Expression Omnibus(GEO), GeneCards, Online Mendelian Inheritance in Man(OMIM), and Therapeutic Target Database(TTD). The common targets shared by Liurui Capsules and EAU were identified, and the protein-protein interaction(PPI) network was established via STRING. Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were conducted via g: Profiler. The rat model of EAU was induced by interphotoreceptor retinoid-binding protein(IRBP) and treated with Liurui Capsules. The inflammatory response of anterior segment and the pathological morphology of retina were observed. The mRNA and protein levels of delta-like ligand 4(DLL4), Notch1, interleukin-17(IL-17), and tumor necrosis factor-alpha(TNF-α) were determined by real-time quantitative PCR(q-PCR) and Western blot, respectively. The network pharmacology analysis predicted 51 common targets of Liurui Capsules and EAU, which were mainly involved in IL-17, TNF, and nuclear factor-kappa B(NF-κB) signaling pathways, as well as liposome receptors and other biological processes. Compared with the control group, the modeling of EAU caused inflammatory changes in the anterior segment and retina and up-regulated mRNA and protein levels of DLL4, Notch1, IL-17, and TNF-α in ocular tissue. Compared with the model group, Liurui Capsules reduced the inflammatory reaction of anterior segment and retina and down-regulated the mRNA and protein levels of DLL4, Notch1, IL-17, and TNF-α. Liurui Capsules can down-regulate the expression of the proteins involved in DLL4/Notch1/IL-17 signaling pathway in ocular tissue and alleviate the ocular inflammation, which may be one of the mechanisms of Liurui Capsules in the treatment of EAU.

[Response Characteristics of Algal Chlorophyll-a to Nitrogen, Phosphorus and Water Temperature in Lake Erhai Based on Quantile Regression].

Based on water quality monitoring data of Lake Erhai from 1990 to 2013,the yearly and seasonally response characteristics of algal chlorophyll-a to total nitrogen (TN),total phosphorus (TP) and water temperature was investigated using quantile regression method.It indicated that water temperature was always the primary limiting factor of algal biomass indicated by chlorophyll a (Chl-a) across recent 24 years,but its limiting effect on algal Chl-a content was kept descending sharply and replaced much by nutrients (e.g.P,N) with the increasing eutrophic level.Especially the slope values of TP on Chl-a were kept ascending from -0.3 to 0.8.Since 2002,the positive effect of TN on Chl-a presented slow descending tendency,and P became the most important limiting nutrient factor of algal growth.According to the seasonal variation analysis,water temperature and N had dominant effects on algal Chl-a in spring and autumn,but they were replaced by P in the case of Chl-a>8 mg·m-3 in spring.In summer,N and P had strong interactive impacts on the algal growth,and the positive effect of P was stronger than that of N in the case of Chl-a>3 mg·m-3.In winter,algal biomass was co-limited by temperature and P.It suggested that P reduction is the prior choice of eutrophication control of Lake Erhai,and simultaneously reducing N load is necessary in the background of global warming.In addition,so far a emergency controlling measure should be taken to monitor algal blooming due to sharp ascending of water temperature within a few days in spring and autumn.

Two New Phenanthrene Glucosides from Cremastra appendiculata and their Cytotoxic Activities.

Two new phenanthrene glucosides, named 2'-hydroxy-4,4',7'-trimethoxy-1, l'-biphenanthrene-2,7-di-O-ß-D-glucoside (1) and 1-(4-hydroxybenzyl)-4-methoxy-2,7-dihydroxy-phenanthrene-8-O-ß-D-glucoside (2), together with three known compounds were isolated from the tubers of Cremastra appendiculata (D.Don) Makino. Their structures were elucidated on the basis of spectral data. Compounds 1-4 showed moderate cytotoxic activity and compound 5 showed weak cytotoxic activity against the two cell lines tested. This is the first reported occurrence of an unusual biphenanthrene glucoside in this plant.

[Impacts on repeated common cold for the adults with different constitutions treated by acupoint application in the dog days and the three nine-day periods after the winter solstice].

OBJECTIVE: To observe the impacts on repeated common cold for the adults with different constitutions treated by acupoint application in the dog days (the three periods of the hottest days) and the three nine-day periods after the winter solstice (the three periods of the coldest days). METHODS: One hundred and fifty-two cases of repeated common cold were divided into four zones according to the body constitution. Each zone was sub-divided into a group of the dog days + the three nine-day periods of the coldest days (group A), and a simple group of the dog periods (group B). In both groups, Dazhui (GV 14), Feishu (BL 13), Tiantu (CV 22), Danzhong (CV 17), Zhongfu (LU 1) and Shenshu (BL 23) were selected. In group A, the acupoint application was given on the 1st or 2nd day of the first, second and third periods of the hottest days in 2010, as well as the 1st or 2nd day of the first, second and third periods of the coldest days in 2010 separately. In group B, the acupoint application was only given on the 1st or 2nd day of the first, second and third periods of the hottest days in 2010. The follow-up visit was conducted before the acupoint application in the three periods of the coldest days in 2010 and before the acupoint application in the three periods of the hottest days in 2011. Additionally, the frequency of disease attack and the symptom score in sickness were taken as the observation indices for the efficacy assessment in both groups. RESULTS: (1) In both groups, the attack frequency was reduced obviously in half a year after the three periods of the hottest days for the patients of qi deficiency constitution, yang deficiency constitution and qi stagnation constitution and the clinical symptom score were reduced apparently (all P<0.01), which were superior to those for the patients of phlegm damp constitution (P<0.01, P<0.05). For the patients of phlegm damp constitution, only the clinical symptom score was reduced (P<0.01). (2) In group A, the improvements were received in the attack frequency and the clinical symptom score in half a year after the three periods of the coldest days for the patients of those four constitutions as compared with those before treatment (all P<0.01). In group B, the attack frequency and the clinical symptom score were all back to the level as those before treatment. (3) In group A, in half a year after the three periods of the coldest days, the efficacy for reducing the attack frequency and the improvements in the clinical symptoms were better than those in group B (all P<0.01). CONCLUSION: The acupoint application in the dog days and the three nine-day periods after the winter solstice reduces the attack frequency and relieves the clinical symptoms of common cold for the adults of individual constitutions to different extents. The efficacy can be further improved in the treatment with the acupoint application in the three nine-day periods after the winter solstice half a year after the acupoint application in the dog days.

[Nutrient dynamics over the past 100 years and its restoration baseline in Dianshan Lake].

The restoration of eutrophic lakes requires a good knowledge on the history and baseline of nutrients in the lakes. This work conducted an analysis on 210Pb/137Cs, water content, loss-on-ignition, sedimentary total phosphorus (TP), total nitrogen (TN), total organic carbon (TOC) and diatoms in the four sediment cores from Dianshan Lake (near Shanghai City). Good coherence in palaeoproxies between the cores indicates a relatively stable sedimentary environment. With increasing human impact, diatom communities shifted from oligo-trophic species Cyclotella bodanica, C. ocelata, Achnanthes minutissima, Cocconeis placentula var lineate, Cymbella sp. , Fragilaria pintata, F. brevistrata, F. construens var venter to recent eutrophic species including Cyclostephanos dubias, C. atomus, Stephanodiscus minitulus, S. hantzschi, Aulacoseria alpigena. The epilimnetic TP over the past 100 years reconstructed using an established diatom-TP transfer function matches well with the monitoring TP where exists. Based on the sedimentary nutrient characteristics and diatom-reconstructed nutrient dynamics, we proposed that the nutrient baseline for Dianshan Lake is 50-60 microg x L(-1), 500 mg x kg(-1) and 550 mg x kg(-1) for water TP concentration, sedimentary TP and TN, respectively.

[Regional heterogeneity of lake eutrophication effects in China].

Although biomass of algae (Chl-a) were in variant levels between different lake regions in China under the same nutrients conditions, it demonstrated that efficiencies of TN/TP used by algae had regional differences. In order to clarify the differences, curve estimation in SPSS was used to analyze the linear relationship between Ig Chl-a and Ig TN/lg TP. The slopes of these linear equations were identified as the efficiencies of TN/TP used by algae. The slopes of linear equations from Mengxin Plateau, Yungui Plateau, Northeast Mountain-Plain, lower reach of Yangtze River Plain and North Plain were 1.002, 0.817, 0.761, 0.545, 0.250, orderly. The efficiencies of TN used by algae ranged from the highest to the lowest were lower reach of Yangtze River Plain, Yungui Plateau, North Plain, Northeast Mountain-Plain, Mengxin Plateau, and the slopes of linear equations were 1.401, 1.058, 0.447, 0.239, 0. 099, respectively. Consequently, in Northeast Mountain-Plain, Mengxin Plateau the efficiencies of TP used by algae were higher than those of TN, and in Yangtze River Plain, Yungui Plateau, North Plain, the efficiencies of TN used by algae were higher than those of TP. On the other hand, in order to describe the effects of algae on transparency in different lakes, the relationships between Chl-a and SD were analyzed. The results showed that in Yungui Plateau the effect of algae on transparency was the most obvious as the variation of SD explained by Chl-a was the highest, and Northeast Mountain-Plain, Mengxin Plateau and North Plain followed. However, in lower reach of Yangtze River Plain, the relationship between Chl-a and SD was not significant.

[Observation of the therapeutic effect on COPD of cold phlegm blocking the lung type at stable stage treated with acupoint sticking therapy in different season].

OBJECTIVE: To systematically observe the clinical effect on chronic obstructive pulmonary disease (COPD) at the stable stage, differentiated as cold phlegm blocking the lung type, treated with acupoint sticking therapy during the dog days and the three nine-day periods after the winter solstice so as to propose the latest clinical idea and theoretic evidence for the treatment of COPD. METHODS: One hundred and fifty cases of COPD at stable stage, which were in accordance with the inclusive standard were randomly divided into three groups, named group A (treatment in dog days and the three nine-day periods after the winter solstice), group B (treatment in dog days) and group C (treatment in the three nine-day periods after the winter solstice), 50 cases in each group. The ingredients (Semen Brassicae, Euphoribia Kansui, Asarum, Rhizome Corydalis, Cinnamon, ginger juice) and doses of herbal medicine plaster were same in each group. The herbal plaster was applied to Feishu (BL 13), Shenshu (BL 23), Dazhui (GV 14), Tiantu (CV 22), Danzhong (CV 17) and Zhongfu (LU 1). In group B, the treatment was given once on the 1st day of each dog-day period, totally 3 treatments were included. In group C, the treatment was given once on the 1st day of each nine-day periods after the winter solstice, totally, 3 treatments were involved. In group A, the treatment was given once on the 1st day of each dog-day period and each nine-day periods after the winter solstice separately, totally 6 treatments were required. The therapeutic effect was evaluated in 4 aspects, named comprehensive clinical efficacy, survival quality (the scores for symptoms, activity limitation and influence on daily life), the attach frequency and pulmonary function. RESULTS: The total effective rate was 88.0% (46/50) in group A, which was superior to 76.0% (38/50) in group B and 70.0% (35/50) in group C separately (P < 0.01, P < 0.001). The results of the attack frequency, clinical symptom score and pulmonary function indices after treatment were all improved apparently as compared with those before treatment in each group (all P < 0.01). All the above indices in group A were improved much apparently as compared with the other two groups (P < 0.01, P < 0.001). Except for the level of forced vital capacity (FVC), the results of clinical symptom score and the other pulmonary function indices in group B were all improved significantly as compared with group C (P < 0.05, P < 0.001). CONCLUSION: Acupoint sticking therapy during different season of the year achieves a superior clinical efficacy for the patients with COPD at stable stage. This therapy can reduce the attack frequency and improve the survival quality and pulmonary function for the patients. It is concluded that the efficacy of the treatment in dog days and the three nine-day periods after the winter solstice is superior to simple dog-day treatment and the treatment in the three nine-day periods after the winter solstice, and the efficacy of dog days treatment is better than that in the three nine-day periods after the winter solstice.

[Therapeutic efficacy of modified zigui decoction in treatment of polycystic ovary syndrome of gan-shen yin deficiency syndrome].

OBJECTIVE: To study the therapeutic efficacy of Modified Zigui Decoction (MZD) in treatment of polycystic ovary syndrome of Gan-Shen yin deficiency syndrome. METHODS: 66 polycystic ovary syndrome patients of Gan-Shen yin deficiency syndrome were randomly assigned to the MZD group (Group A) and the Westem medicine group (Group B), 33 patients in each. Patients in Group A orally took MZD, while those in Group B orally took Diane-35. Their menstrual cycle rate, basal body temperature (BBT), the ovarian size, the number of follicles, and changes of endocrine hormones were observed before treatment, the first menstrual cycle, and the sixth menstrual cycle after treatment. RESULTS: (1) The normal rate of one menstrual cycle after stopping taking medicine was 57.58% in Group A and 63.64% in Group B. There was no statistical difference between the two groups (P>0.05). The normal rate of six menstrual cycles after stopping taking medicine was 45. 45% in Group A and 21.21% in Group B. The former was superior to the latter, showing statistical difference (P<0.05). (2) The biphasic BBT rates of one menstrual cycle after stopping taking medicine were somewhat elevated in the two groups, better than before treatment respectively (P<0.01). But there was no statistical difference between the two groups (P>0.05). The biphasic BBT rate of six menstrual cycles after stopping taking medicine was 45.45% in Group A and 18.18% in Group B. The former was superior to the latter, showing statistical difference (P<0.05). (3) The bilateral ovarian volume of one menstrual cycle after stopping taking medicine was obviously reduced in both groups (P<0.01). The bilateral ovarian volume of six menstrual cycles after stopping taking medicine was still more reduced than before treatment in Group A (P<0.01), while it returned to the size of before treatment in Group B (P>0.05). (4) The number of follicles of one menstrual cycle after stopping taking medicine was obviously reduced in both groups (P<0.01). The number of follicles of six menstrual cycles after stopping taking medicine was still reduced in Group A (P<0.01), while it returned to the number before treatment in Group B (P>0.05). (5) The luteinizing hormone (LH), testosterone (T), LH/FSH ratio of one menstrual cycle after stopping taking medicine were obviously reduced in both groups (P<0.01). They were still more reduced six menstrual cycles after stopping taking medicine than before treatment in Group A (P<0.01), while they returned to the levels of before treatment in Group B (P>0.05). CONCLUSIONS: MZD could effectively treat patients with polycystic ovary syndrome of Gan-Shen yin deficiency syndrome. Besides, its long-term efficacy was more stable and lasting.

[KeLuoXin medicated serum extract inhibits high glucose-induced proliferation and phosphorylation of ERK1/2 in rat mesangial cell].

OBJECTIVE: To investigate the effects and mechanism of KeLuoXin (KLX) capsule on rat mesangial cell proliferation induced by high glucose. METHODS: Rat mesangial cells (HBZY-1) were cultured in vitro and stimulated with high glucose(30 mmol/L glucose) for different time after the treatment with or without KLX medicated serum extract, the proliferation was assessed by cell counting Kit-8 (CCK-8) and the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) was detected by Western blot. RESULTS: KLX could significantly inhibit HBZY-1 cell proliferation induced by high glucose (P < 0.05). In addition, KLX could reduce the increase of the phosphorylation level of ERK1/2 (P < 0.01), which was induced by high glucose in HBZY-1 cells. CONCLUSION: KLX can inhibit rat mesangial cell proliferation, and the mechanisms may relate to the interruption of ERK1/2-Mitogen-activated protein kinase (MAPK) pathway.

Response of sodium pump to ouabain challenge in human glioblastoma cells in culture.

Bipolar disorder is a severe psychiatric condition that manifests with abnormalities in ion regulation. Previous studies have suggested that glia may be specifically involved in the pathophysiology of this condition. Since the potent sodium pump inhibitor, ouabain, has been used previously to model the ionic changes of bipolar illness, we investigated its effect of on sodium pump expression and activity in a human glioblastoma cell line. LN229 cells were grown with or without ouabain 10(-7) M for 3 days, and the effect of a therapeutic concentration of lithium was also examined. The mRNA transcription of sodium pump isoforms was determined by reverse transcriptase polymerase chain reaction (RT-PCR), and the protein expression of phosphorylated and non-phosphorylated pump isoforms was semi-quantified utilizing Western blot. Ouabain treatment caused an increase of some 6-fold in alpha1 protein expression and a doubling of alpha1 mRNA. alpha3 protein and alpha2 and alpha3 mRNA more than doubled. Lithium treatment alone had no effect, but lithium co-administered with ouabain normalized Na pump protein and mRNA expression for alpha1 and 2, but not alpha3. These results suggest that disturbance of ion regulation induces changes in glial cell sodium regulatory systems which are normalized by lithium treatment.

[Environmental impacts on periphyton assemblage in Shanghai's main urban rivers, China].

A monthly monitoring program of periphyton was conducted on two main urban rivers of Shanghai from April 2005 to March 2006. Parameters of periphyton assemblage and main environmental variations, including physical habitation and hydrological factors, were analyzed by multivariate analyses (cluster analysis, indicator species analysis and canonical correspondence analysis). It showed that a remarkable spatial-temporal variation pattern was observed for periphyton assemblage: taxa numbers and density were abundant at upstream in warm weathers, while diversity decreased significantly at downstream in cold seasons (p < 0.05). The results also indicated that water quality, watershed function zone and nutrients (i.e. nitrogen and phosphorus) were the main environmental factors influencing periphyton species composition. The findings further suggested 12 periphyton species, including Navicula cincta, Chroococcus minutus and Fragilaria capucina, had significant indicator values for water quality and were highly sensitive to the concentrations of nitrogen and phosphorus, and these species could serve as a good bio-indicator of urban watersheds.

[Correlation between sex hormone and insulin and various TCM syndrome types in patients with polycystic ovarian syndrome].

OBJECTIVE: To explore the relationships of sex hormone and insulin with various TCM syndrome types of polycystic ovarian syndrome (PCOS) and their significance. METHODS: Levels of testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), estradiol (E2), and insulin (INS) in 120 patients with PCOS were determined respectively, among them, 30 patients were classified by TCM syndrome differentiation as Shen-yin deficiency type (SYiD), 30 as Shen-yang deficiency type (SYaD), 30 as Pi-yang deficiency type (PYD) and 30 as Gan-stagnancy transformed heat type (GSH). RESULTS: It was shown that T level in SYiD, serum LH and LH/FSH ratio in SYaD, PRL in GSH and INS in PYD patients were higher as compared with those in patients of other three types (P < 0.05 or P < 0.01), except for the above-mentioned, the differences in all the paired comparisons of all the indexes between TCM types were insignificant. CONCLUSION: Elevated level of T, LH and LH/FSH ratio, PRL and INS are correlated with the TCM syndrome type of SYiD, SYaD, GSH and PYD in patients with PCOS respectively, and being the clinical characteristics of the corresponding syndromes.

[Identification of Epimedium Koreanum Nakai by Fourier transfer infrared spectroscopy].

The herbal plant Epimedium Koreanum Nakai is a native herb growing in the Changbai Mountain region, Jilin province of China, and is used as a tonic, anti-rheumatic and aphrodisiac in traditional Chinese medicine. The analytical method existing for the identification of Epimedium Koreanum Nakai is time-consuming, and can not be used for fast, in situ and nondestructive analysis. Fourier transform infrared spectroscopy was used to identify wilding and planting Epimedium Koreanum Nakai. The spectra of 18 samples were analyzed. They show some differences in the intensity and the shape of absorption peaks in the range of 1,800-1,200 cm(-1). The absorption intensity of the spectra changed regularly with the producing area. So FTIR spectra could be used for the identification of Epimedium koreanum Nakai.

[Effect of aluminum trichloride on dissociated Ca2+ in Hippocampus neuron cell as well as learning and memory].

OBJECTIVE: To investigate the effect of aluminum chloride on dissociated Ca(2+) in hippocampus neuron cells in mice and the relationship to the learning and memory. METHODS: Male ICR mice in the three intoxicated groups were administered with the double distilled water containing AlCl(3) (10, 50, 300 mg.kg(-1).d(-1)) while those in the control group were administered with the double distilled water for 100 days. The methods of behavior toxicology such as Morris swim maze were used for studying the effect of aluminum chloride on the changes of learning and memory in mice. With calcium sensitive fluorescence indicator Fura-2 as the fluorescent probe, the influence of the subchronic exposure to Al on the dissociated Ca(2+) in hippocampus neuron cells was observed. RESULTS: The dissociated Ca(2+) in hippocampus neuron cells in the middle dosage group and the high dosage group [(412.25 +/- 53.20), (467.37 +/- 32.85) times] was lower than those in the control group [(293.91 +/- 32.21) times] respectively (P < 0.01), and correlated positively with the dose and dissociated Ca(2+) (r = 0.861, P < 0.01). Compared with the control group, the latent period was lengthened (P < 0.05) in the middle dosage and the high dosage group. CONCLUSION: The subchronic exposure to AlCl(3) in mice affects the dissociated Ca(2+) in hippocampus neuron cells. The increase of dissociated Ca(2+) in hippocampus neuron cells may be correlated with the disfunction of cognition in the aluminium intoxicated mice.

Individualization of tiger by using microsatellites.

In investigating criminal cases of poaching and smuggling involving tigers (Panthera tigris), the number of tiger individuals involved is critical for determining the penalty. Morphological methodologies do not often work because tiger parts do not possess the distinctive characteristics of the individual. Microsatellite DNAs have been proved a reliable marker for the individualization of animals. Seven microsatellite loci derived from domestic cat (Felis catus) were selected to individualize tigers, namely F41, F42, F146, Fca304, Fca391, Fca441 and Fca453. A reference population containing 37 unrelated tigers were used to investigate alleles, allelic frequencies, genotypes and genotype frequencies of each locus. Consequently, the data was used to assess the validity of the combination of seven loci for tiger individualization. All loci were polymorphic and easy to amplify. Three out of the seven loci were significantly departure from the Hardy-Weinberg Equilibrium (P < 0.05). Cumulative discrimination power (DP) calculated with observed genotype frequencies was 0.99999789. Match probability of an individual in the reference population with a random individual in seven loci ranged from 7.34 x 10(-9) to 2.77 x 10(-5). This suggests that combining the seven microsatellite loci provides desirable power to individualize tigers. The combination of seven loci was applied to a case of tiger bone smuggling. Genotypes of all samples were identical in all seven loci, and the P(M) of the evidence samples in the seven loci hit 5.63 x 10(-7), provided evidence that the bones belong to a single tiger.