[Transcriptome profiling of Saposhnikovia divaricata growing for different years and mining of key genes in active ingredient biosynthesis].

Saposhnikovia divaricata is a commonly used bulk medicinal plant. To explore the key enzyme genes and their expression in the biosynthesis of chromone and coumarin, the key active components, we carried out transcriptome sequencing(Illumina HiSeq) and bioinformatics analysis for the 1-year-old(S1) and 2-year-old(S2) plants of S. divaricata. A total of 40.8 Gb data was obtained. After the sequence assembly via Trinity, 110 732 transcripts and 86 233 unigenes were obtained, which were aligned and annotated with NR, Swiss-Prot, GO, KEGG, and PFAM. Daucus carota and S. divaricata had the highest sequence homology. KEGG pathway enrichment showed that the differentially expressed genes were mainly enriched in plant hormone signal transduction, phenylpropanoid biosynthesis, and flavonoid biosynthesis pathways. A total of 27 differentially expressed unigenes, including 13 enzyme genes, were identified in the pathways related to the synthesis of active ingredients in S. divaricata. Compared with S1 plant, S2 plant showed up-regulated expression of PAL, BGL, C4H, 4CL, CYP98A, CSE, REF, and CCoAOMT and down-regulated expression of CHS, CAD, and COMT. HCT and POD had both up-regulated and down-regulated unigenes. Among them, PAL, C4H, 4CL, BGL, and CHS can be used as candidate genes for the synthesis of the active ingredients in S. divaricata. The four key enzyme genes were verified by RT-qPCR, which showed the results consistent with transcriptome sequencing. This study enriches the genetic information of S. divaricata and provides support for the identification of candidate genes in the biosynthesis of secondary metabolites.

[Clinical characteristics of Yinhua Miyanling Pian users based on real world study].

Clinical characteristics of Yinhua Miyanling Pian user group were analyzed based on real world hospital information system (HIS)database. The information was collected from the HIS in sixteen hospitals of grade Ⅲ-A. Normalizing the data and descriptive analysis was performed. Among the 5 312 cases, female patients(63.67%) were more than male patients (36.33%). The median age was 49 years old. The age of 18-65 accounted for 74.52%.The patient was admitted to the hospital's department of obstetrics and gynecology(28.71%) and urology(28.43%). The median single dose is 2 g, accounted for 49.55%. The median daily dose is 6 g. 88.80% of patients were treated for less than 7 d. The median hospitalization were 12 d, most were 7-14 d accounted for 41.70%. Most hospitalization expenses payment by medicare, accounted for 80.22%. The median hospitalization expenses was 12 211.47 RMB. Most patients with benign tumor(27.36%) and malignant tumor(15.56%), next is the obstruction of urinary tract(15.49%) and urinary calculi(10.52%). The most common syndromes were damp heat syndrome(32.46%), liver and kidney deficiency syndrome(15.33%) and splenasthenic fluid-retention syndrome(15.01%). Clinical use is combined with antibiotics, as well as traditional Chinese medicine heat clearing agents, tonic drugs and so on. Finally, 44.22% were cured and 51.05% were better. Most of the drug users were adults, and mostly were female. Most with the tumor, urinary tract obstruction or stones. TCM syndrome is mainly characterized by dampness and heat, deficiency of liver and kidney, and dampness of spleen. In clinical practice, it was mainly combined with Western medicines, and Chinese medicines were also a-vailable. And most are combined with antibiotics.The medication basically conforms to the drug instruction. Based on the results of the real world HIS，Yinhua Miyanling Pian could provide theidea and reference for regulating the medication in adolescent patients.

[Effect of Modified Leweiyin Recipe on SGC-7901 Human Gastric Cancer Cells from Crosstalk between NF-κB and STAT3].

Objective To observe the effect of Modified Leweiyin Recipe (MLR) on nuclear factor kappa B (NF-κB) and crosstalk between signal transducers and activators of transcription 3 (STAT3) in SGC-7901 human gastric cancer cells. Methods SGC-7901 human gastric cancer cell strain was taken as subjects. The vectors of NF-κB (ReIA/p65)-PECFP and STAT3-PEYFP were constructed and transfected in SGC-7901 human gastric cancer cells. Cells were then intervened by MLR after stimulated by LPS. The crosstalk between NF-κB and STAT3 in cells was detected using fluorescence resonance energy transfer and co-immunoprecipitation. Results After LPS stimulation, the crosstalk between NF-κB and STAT3 was enhanced. But it was significantly weakened after MLR intervention. Conclusion MLR could treat precancerous lesions of gastric cancer and prevent the occurrence of gastric cancer possibly by blocking the crosstalk between NF-κB and STAT3.

Effect of glutamine enriched nutrition support on surgical patients with gastrointestinal tumor: a meta-analysis of randomized controlled trials.

BACKGROUND: Associations between glutamine (Gln) enriched nutrition support and surgical patients with gastrointestinal (GI) tumor remain controversy. The purpose of this meta-analysis was to assess the effect of Gln enriched nutrition support on surgical patients with GI tumor in term of relevant biochemical indices, immune indices, and clinical outcomes. METHODS: Six databases were systematically searched to find eligible randomized controlled trials (RCTs) from 1966 to May 2014. When estimated the analysis indexes, the relative risk (RR) was used as the effect size of the categorical variable, while the weighted mean difference (MD) was used as the effect size of a continuous variable. Meta-analysis was conducted with Rev Man 5.2. RESULTS: Thirteen RCTs, involving 1034 patients, were included in the meta-analysis. The analysis showed that Gln enriched nutrition support was more effective in increasing serum albumin (MD: 0.10; 95% confidence interval [CI]: 0.02-0.18; P < 0.05), serum prealbumin (MD: 1.98; 95% CI: 1.40-2.55; P < 0.05) and serum transferring (MD: 0.35; 95% CI: 0.12-0.57; P < 0.05), concentration of IgG (MD: 1.26; 95% CI: 0.90-1.63; P < 0.05), IgM (MD: 0.18; 95% CI: 0.11-0.25; P < 0.05), IgA (MD: 0.22; 95% CI: 0.10-0.33; P < 0.05), CD3 + (MD: 3.71; 95% CI: 2.57-4.85; P < 0.05) and CD4/CD8 ratio (MD: 0.27; 95% CI: 0.12-0.42; P < 0.05). Meanwhile, it was more significant in decreasing the incidence of infectious complications (RR: 0.67; 95% CI: 0.50-0.90; P < 0.05) and shortening the length of hospital stay (MD: -1.72; 95% CI: -3.31--0.13; P < 0.05). CONCLUSIONS: Glutamine enriched nutrition support was superior in improving immune function, reducing the incidence of infectious complications and shortening the length of hospital stay, playing an important role in the rehabilitation of surgical GI cancer patients.

[Temporal and Spatial Distribution of Nutrients in Daning River Sediments and Their Correlations with Chlorophyll in the Three Gorges Reservoir Area].

In order to understand the temporal and spatial distribution of nutrients in Daning river sediments and to investigate the relationship between nutrients and algal cells, samples were selected from 4 typical sampling sites, i. e. Caizi Dam, Baishui River, Shuanglong and Dachang. Vertical gravity mud samplers were used to collect the sediments from different layers within 20 cm in increment of 2 cm. Vertical distributions of TN, NH4(+)-N, NO3(-) -N, NO2(-) -N, TP, inorganic P and organic P in sediments of different sampling sites were investigated. And so was the vertical distribution of chlorophyll. Moreover, the correlation between chlorophyll and the nutrients was analyzed. Results showed that the contents of TN were the highest at the depth of 0.0-2.0 cm in the Caizi Dam sediments, and at the depth of 2.0-4.0 cm in the Baishui River sediments. The highest NH4(+) -N content occurred at 2.0-4.0 cm in the Caizi Dam sediments and at 4.0-6.0 cm in the Baishui River in January and February. As to NO3(-) -N and NO2(-) -N contents, they were the highest at 2.0-4.0 cm in Caizi Dam sediments except March. For NH4(+) -N, NO3(-) -N and NO2(-) -N, there was no significant difference under 4.0 cm sediments at the same sampling site. The distribution of TP was increasing from 0.0 to 6.0 cm in Baizi River sediments. But in Caizi Dam sediments TP and inorganic P contents in 0.0-2.0 cm were higher than those of others layers; TP and inorganic P in all different layers of Caizi Dam sediments were greater than those of corresponding layers in other sampling sites' sediments. The contents of organic P at Caizi Dam and Dachang were higher than those of Baishui River and Shuanglong, while the difference of organic P contents was not significant in different sediments layers at the same sampling site. The chlorophyll a contents in Dachang were the highest at every layer comparing to the corresponding layers of others, followed by Baishui River, Caizi Dam, and Shuanglong. Only one significantly positive correlation was observed between the content of inorganic P and chlorophyll a in Dachang site in January, with the correlation coefficient was 0.87. The correlation results implied that nutrients in sediments were not the main factors affecting the distribution of algal cells.

[Effects of grain-sized moxibustion on learning and memory ability and amyloid deposition of transgenic Alzheimer's disease mice].

OBJECTIVE: To observe the effect of grain-sized moxibustion at "Xinshu" (BL 15) and "Shenshu" (BL 23) on memory-learning ability and amyloid deposition in transgenic Alzheimer's disease (AD) mice. METHODS: seventeen amyloid precursor protein (APP)/presenilin (PS)1 (APP+/PS 1+) double transgenic 6799 mice aged 3-4 weeks were randomly divided into model group (n = 9) and moxibustion group (n = 8). Nine wide-type (C 57 BL/6 J) female mice were used as the normal control group. Moxibustion (ignited grain-sized moxa cone) was applied to bilateral "Xinshu" (BL 15) and "Shenshu" (BL 23) for about 30 s, once a day for 9 courses (10 days constitute a therapeutic course, with 2 days' break between every two courses). Morris water maze tests were performed to detect the mice's learning-memory ability. The alterations of beta-amyloid deposition (number of the positive plaques) in the cerebral cortex and hippocampus were detected by using an imaging analysis system following Congo red staining of the cerebral tissue sections. RESULTS: Compared with the normal group, the average escape latency of place navigation tests was significantly increased (P < 0.05), and the target-platform crossing times and percentage of dwell time in the target quadrant of the spatial exploring tests were notably decreased in the model group (P < 0.05). After moxibustion intervention, the escape latency was remarkably shortened, and the target-platform crossing times and dwell time percentage were obviously increased in the moxibustion group in comparison with the model group (P < 0.05), suggesting an improvement of the learning-memory ability after moxibustion. Results of Congo red staining of the cerebral tissue showed that there were many irregular, uneven staining positive plaques in the cerebral cortex and hippocampus of AD mice in the model group. Compared with the model group, the positive plaque numbers in both cerebral cortex and hippocampus were considerably reduced in the moxibustion group (P < 0.05). CONCLUSION: Grain-sized moxibustion at "Xinshu" (BL 15) and "Shenshu" (BL 23) can ameliorate learning-memory ability and restrain the formation of amyloid deposition in AD mice.

Strong inhibition of celastrol towards UDP-glucuronosyl transferase (UGT) 1A6 and 2B7 indicating potential risk of UGT-based herb-drug interaction.

Celastrol, a quinone methide triterpene isolated from Tripterygium wilfordii Hook F., has various biochemical and pharmacological activities, and is now being developed as a promising anti-tumor agent. Inhibitory activity of compounds towards UDP-glucuronosyltransferase (UGT) is an important cause of clinical drug-drug interactions and herb-drug interactions. The aim of the present study is to investigate the inhibition of celastrol towards two important UDP-glucuronosyltransferase (UGT) isoforms UGT1A6 and UGT2B7. Recombinant UGT isoforms and non-specific substrate 4-methylumbelliferone (4-MU) were used. The results showed that celastrol strongly inhibited the UGT1A6 and 2B7-mediated 4-MU glucuronidation reaction, with 0.9 ± 0.1% and 1.8 ± 0.2% residual 4-MU glucuronidation activity at 100 µM of celastrol, respectively. Furthermore, inhibition kinetic study (Dixon plot and Lineweaver-Burk plot) demonstrated that celastrol noncompetitively inhibited the UGT1A1-mediated 4-MU glucuronidation, and competitively inhibited UGT2B7-catalyzed 4-MU glucuronidation. The inhibition kinetic parameters (Ki) were calculated to be 0.49 µM and 0.045 µM for UGT1A6 and UGT2B7, respectively. At the therapeutic concentration of celastrol for anti-tumor utilization, the possibility of celastrol-drug interaction and celastrol-containing herbs-drug interaction were strongly indicated. However, given the complicated nature of herbs, these results should be viewed with more caution.

[Vertical migration of algal cells in the Daning River Bay of the Three Gorges Reservoir].

To support the basic data for forecast of algal blooms, circadian vertical migration experiment was carried out in the Daning River Bay of the Three Gorges Reservoir in July, 2011. The results were as follows: in this period, different algal species were found in the Daning River Bay, including cyanobacteria, green algae, dinoflagellates and diatoms etc; the distribution of algal cells was uneven, 72.5%-76.2% of algal cells aggregated at 0.5-4.0 m water depths from 10:00 AM to 10:00 AM next day, but 7.5%-16.3% of algal cells aggregated at 0-0.5 m. Morisita's indexes (MI) of algal cells were from 1.41 to 1.97 in day time, and from 1.17 to 1.55 at night. Morisita's indexes of chlorophyll a(Chla)were from 1.31 to 1.59 in day time, and from 1.17 to 1.39 at night. The vertical migration also occurred at 0.5-4.0 m water depths. The algal density was not significantly related with the concentrations of nutrient except that of dissolved total phosphorus (DTP) (r = 0.89). The algal density was also influenced by water temperature (WT), pH and specific conductance (SPC), and the correlation coefficients were 0.96, 0.97 and -0.99, respectively.

Association of apolipoprotein E 4 polymorphism with cerebral infarction in Chinese Han population.

AIM: To study the association between APOE polymorphisms and cerebral infarction through a case-control study among the Chinese Han population. METHODS: First-ever cerebral infarction patients (n=226) whose ages ranged from 40 to 60 years old were recruited from Department of Neurology, Zhongshan Hospital, Shanghai, and Zhejiang Chinese Traditional Medicine Hospital, Zhejiang, China. Unrelated healthy controls (n=201) were selected from the general population in the same area with similar age and sex distribution. APOE was amplified by one-stage PCR using the forward primer: 5'-GGC ACG GCT GTC CAA GGA GCT-3' and reverse primer: 5'-GAT GGC GCT GAG GCC GCG CT-3'. The PCR product was digested directly with 5 U of CfoI and separated by a 20 % polyacrylamide (acrylamide: bis-acrylamide=29:1) nondenaturing gel. RESULTS: Both cerebral infarction patient and control groups were in Hardy-Weinberg equilibrium. The allele frequency of APOE*2, APOE*3, and APOE*4 was 4.6 %, 81.9 %, and 13.5 % respectively in the patients with cerebral infarction; 5.7 %, 87.3 %, and 7.0 % respectively in the healthy control group. Compared with APOE3/3 subjects, APOE4/4 carriers had a 2.1-fold risk of cerebral infarction (odds ratio 2.1, 95 % confidence limits 1.3 to 3.4). The allele frequency of APOE*4 in the cerebral infarction patient group was significantly higher than that in the control group (13.5 % vs 7.0 %; P=0.002). CONCLUSION: APOE 4 is a risk factor for cerebral infarction among the Chinese Han population.