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An ultra performance liquid chromatography (UPLC) method was established and validated to simultaneously determine the contents of six aconitum alkaloids in mother, daughter and fibrous roots of 19 batches of Aconitum carmichaelii from Sichuan province. The separation of the six alkaloids was achieved on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 microm) column at 40 degrees C with a mobile phase consisting of acetonitrile in 30 mmol x L(-1) ammonium acetate buffer solution (adjusted to pH 10.0 with aqueous ammonia) in gradient mode. The data and plots showed that the six aconitum alkaloids have different distributions. Four aconitum alkaloids were almost same in mother and daughter root except benzoylmesaconine and mesaconitine, while the fibrous root differed from the other two roots. The comparisons of significant differences of six aconitum alkaloids between the mother and daughter roots definitely demonstrated that benzoylmesaconine and mesaconitine were the representative components. The 38 detecting samples were classified as two clusters by hierarchical clustering analysis (HCA) and principle component analysis (PCA), the results indicated that the mother root was different from the daughter root on chemical material basis. The study might contribute to the reasonable clinical application of A. carmichaelii.

Objectives: To investigate the effects of plasma from the patients with preeclampsia on proliferation and apoptosis of human umbilical vein endothelial cells (HUVEC), and to explore the relationship between cell damage and lysophosphatidic acid (LPA) receptors., Methods: Sixty patients with preeclampsia were recruited from October 2011 to June 2012 in the First Affiliated Hospital of Zhengzhou University. Among them, thirty cases were defined as the mild preeclampsia group and thirty cases were defined as the severe preeclampsia group. The other thirty healthy pregnant women were recruited in the healthy pregnant women group. The levels of plasma LPA in the three groups were measured. The HUVEC were cultured in vitro with plasma from the three groups, and a blank control group was set up as well. Proliferation and apoptosis of HUVEC were measured by MTT assay and flow cytometry. Immunohistochemistry of biotin streptomyces protein peroxidase (SP) method was used to measure the protein expression level of Edg 2, 4, 7., Results: (1) The plasma LPA levels in the healthy pregnant woman group, mild preeclampsia group and severe preeclampsia group were (3.38 ± 2.08) µmol/L, (6.12 ± 0.22) µmol/L, (9.10 ± 0.17) µmol/L, respectively. The plasma levels of LPA in patients with preeclampsia were significantly higher than that in the healthy pregnant women (P < 0.01). (2) The proliferation rate of HUVEC in the mild and severe preeclampsia groups [(65.2 ± 2.7)% and (51.9 ± 2.8)%] were significantly lower than that in the healthy pregnant women group and the control group [(84.3 ± 3.1)% and (100.0 ± 0.0)%, P < 0.01]. (3) The early apoptosis rate, middle-late apoptosis rate and total apoptosis rate of HUVEC in the mild and severe preeclampsia groups [total apoptosis rate were (30.4 ± 2.0)% and (43.4 ± 2.5)%] were significantly higher than those in the healthy pregnant women group and the control group [total apoptosis rate were (18.6 ± 1.6)% and (8.0 ± 1.5)%, P < 0.01]. (4) The expression positive rates of Edg 2, 4, 7 proteins in the four groups were as following: mild preeclampsia group 83%, 80% and 73%; severe preeclampsia group 97%, 93% and 90%; healthy pregnant women group 40%, 40% and 37%, and the control group 10%, 10% and 7% respectively. The positive rates of HUVEC in the mild and severe preeclampsia groups were significantly higher than those in the healthy pregnant women group and the control group (P < 0.01)., Conclusions: The plasma of patients with preeclampsia could inhibit proliferation and promote apoptosis of HUVEC, and induce the expression of Edg 2, 4, 7 proteins. It suggested that the increase of lysophosphatidic acid in plasma could be one of the reasons of endothelial cell damage in patients with preeclampsia.

A prospective multicenter clinical trial was conducted to compare the beneficial effects of a Chinese herbal medicine formula Jiangzhuoqinggan (JZQG) and western antihypertension drug irbesartan. JZQG is mainly composed of rhubarb, coptis, cassia, and uncaria. A total of 240 patients with mild to moderate hypertension were enrolled in the trial. Patients were assigned into two groups after screening: JZQG group and the irbesartan group. After four weeks of treatment, we compared the changes in routine blood pressure, 24 h ambulatory blood pressure, and waist circumference. There was a significant reduction in systolic blood pressure and diastolic blood pressure in the JZQG group (both p < 0.01). There were no significant differences between the reduction of systolic and diastolic blood pressures in the two treatment groups. From the 24 h ambulatory blood pressure measurement, the JZQG group showed a greater reduction in both systolic and diastolic blood pressures (in both daytime and nighttime) than the irbesartan group. Furthermore, there was a significant difference in waist circumference in the JZQG group (1.51 cm reduction; P < 0.05) but not the irbesartan group (0.42 cm). Thus, the JZQG formula may have therapeutic value in patients with both hypertension and metabolic syndrome.

Objective: To analyze ginsenosides composition in wild ginseng leaves with different growth years., Method: The analysis was performed on Acquity UPLC BEH Shield RP18 (2.1 mm x 100 mm, 1.7 microm) column, the mobile phase was acetonitrile-0.05% formic acid solution in gradient elution mode. The detection wavelength was at 203 nm. The flow rate was 0.4 mL x min(-1) and column temperature was set at 30 degrees C., Result: Thirteen ginsenosides were determined by the established UPLC method. In 5-17th growth year ginseng leaf samples cultivated simulating wild conditions, the contents of ginsenosides in the 14th year have the highest content., Conclusion: The established method is simple, accurate and reliable, can be used in ginsenosides determination and fingerprint research of Panax crude drug. The result provides reliable data for the accumulation of ginsenosides and sustainable utilization of ginseng resources.

Objective: To observe the ophthalmic artery (OA), central retinal artery (CRA) and posterior ciliary artery (PCA) blood flow and the changes of eye vascular auto-regulation in patients with obstructive sleep apnea-hypopnea syndrome (OSAS)., Methods: It was a case-control study. Fifteen health overweight male adult (as normal control) and 42 patients with OSAS were randomly selected from examination center and sleep detection center, respectively. The OSAS patients were divided into mild (14 patients) and moderate and severe (28 patients) groups based on the apnea-hypopnea index (AHI). All subjects filled the sleep questionnaire and carried out polysomnogram monitoring all night for at least 7 hours. Fasting peripheral venous blood was collected at 7 AM on next day. The end-tidal CO(2) (ETCO(2)), intraocular pressure and color doppler sonography were examined next day to record the data in the inspection process before and after Mueller maneuver. Doppler ultrasound measurement of ocular blood flow diameter and blood flow velocity values were described in the median (max, min) and compared with Kruskal-Wallis test. And then two groups were compared with Bonferroni t test. Ocular blood flow velocity of patients with OSAS and PSG monitoring indicators were analyzed using partial correlation analysis., Results: OA inner diameter in moderate and severe OSAS group [0.08 (0.15, 0.06) cm] was lower than that in healthy control [0.15 (0.26, 0.11) cm] and the difference was statistically significant (P = 0.000). PCA inner diameter in moderate and severe OSAS group [0.10 (0.13, 0.07) cm] were higher than that in healthy controls [0.05 (0.09, 0.04) cm]. CRA peak systolic velocity (PSV) in moderate and severe OSAS group [16.50 (19.40, 13.10) cm/s] was greater than that in healthy controls [11.30 (16.70, 8.20) cm/s]. The differences between these two groups were statistically significant (PCA inner diameter: P = 0.000, CRA-PSV: P = 0.001). The difference of CRA end diastolic velocity (EDV) between the moderate and severe group [8.90 (9.90, 5.10) cm/s], mild group [7.00 (8.30, 4.50) cm/s] and healthy control group [5.50 (7.40, 3.40) cm/s] was statistically significant (χ(2) = 14.45, P < 0.05). PCA-PSV [32.50 (43.10, 19.10) cm/s] and PCA-EDV [12.80 (15.20, 5.70) cm/s] in the moderate and severe group were higher than those in healthy control group [22.60 (32.20, 12.40) cm/s] and [7.20 (11.20, 3.90) cm/s], as well as those in the mild group [24.00 (30.70, 13.30) cm/s] and [8.00 (9.90, 3.90) cm/s]. These differences were statistically significant (PCA-PSV: P = 0.000, 0.002; PCA-EDV: P = 0.000, 0.001). The diameter of OA and PCA correlated negatively with ETCO(2) (r = -0.41, -0.34; P < 0.05); CRA-PSV was correlated with SaO2 min (r = -0.37, P < 0.05). CRA-EDV was correlated with ETCO(2) and SaO2 mean (r = 0.57, -0.39; P < 0.05). PCA-PSV was correlated with SaO2 min and MAI (r = -0.34, 0.56; P < 0.05). PCA-EDV was correlated SaO2 min and MAI (r = -0.29, 0.61; P < 0.05)., Conclusions: The diameter and blood flow of OA, PCA and CRA change in OSAS patients. Compared with non-OSAS patients, the autoregulation function of PCA and CRA is weakened in OSAS patients.

Objective: To study the effect of quercetin on the proliferation of neural stem cells in the subventricular zone (SVZ) of rats after focal cerebral ischemia., Methods: An adult rat model of middle cerebral artery occlusion (MCAO) model was established by placement of an intraluminal filament at the origin of the MCA. Quercetin was administered intraperitoneally in the rats at a dose of 50 mg/kg every 3 days starting at 6 h after MCAO, and BrdU (50 mg/kg daily) was also injected intraperitoneally starting at 4 h after MCAO. BrdU-positive cells in the SVZ were counted at 7, 14 and 21 days after MCAO., Results: Compared with the sham-operated group, the rats in the ischemic model group showed significantly increased BrdU-positive cells in the ipsilateral SVZ 7 days after MCAO, reaching the peak level on day 14 and beginning to decrease on day 21 (P<0.05). The number of ipsilateral BrdU-positive cells in quercetin group was significantly greater than that in the model group on days 7, 14 and 21 (P<0.05), and maintained the high level on day 21., Conclusion: Quercetin can maintain a high level of neural stem cell proliferation in the SVZ after focal cerebral ischemia in adult rats.

Objective: To study the chemical constituents of roots of Rumex patientia., Methods: The compounds were separated and purified by silica column chromatopraphy, Sephadex LH-20 and identified by several spectroscopic methods., Results: Seven constituents were identified as nepodin (1), nepodin-8-O-beta-D-glucopyranoside (2), chrysophanol-8-O-beta-D-glucopyranoside (3), emodin-6-O-beta-D-glucopyranoside (4), emodin-8-O-beta-D-glucopyranoside (5), 1, 3, 5-trihydroxy-7-methylanthraquinone (6), physcion-8-O-beta-D-glucopyranoside (7), respectively., Conclusion: Compound 1 is isolated from this plant for the first time and compound 6 is isolated from Rumex L. for the first time.

Next generation sequencing has already been used for genomic analysis of microorganis, human being, animals, and plants. Sample preparation is prerequisite and most important for large-scale sequencing. There are two major interferences for large-scale sequencing, polyA and abundant genes' concealment for rare genes. In order to solve these problems, we used total RNA extracted from violaceae leaves to produce double stranded cDNA. DSN nuclease was used to treat the ds cDNA prior to removing the polyA. Randomly sequencing 100 clones of the treated cDNA showed that there were 94 independent clones in the treated sample, and the sequences did not contained polyA. However, only 62 independent clones were found in the untreated sample, and 15 of the sequencing files were affected by polyA. By randomly sequencing of the treated cDNA, we also found two clones encoded two interested genes. We failed to isolate these genes although the protein mass peaks of them had been found in the MALDI-TOF trace. Furthermore, we designed primers from two known genes with different expression abundances. The PCR yields were approaching similar using the treated cDNAs as templates. These results showed that, removal of the polyA and enrichment of rare genes with DSN can meet the requirements of large-scale sequencing and discovery of new genes.

Objective: To investigate the protective effect of mild hypothermia on rat astrocytes with traumatic or ischemic injury., Methods: Rat astrocytes in primary culture were subjected to scratching or hypoxic injury and exposed to normothermia (37 degrees celsius;) or hypothermia (34 or 32 degrees celsius;) for 24 h. The morphology of the astrocytes was evaluated by live/dead staining, and the cell injury was measured by lactate dehydrogenase (LDH) release assay., Results: As the temperature reduced the LDH release rate from the cells in hypoxic group decreased significantly, to (11.48 - or + 1.53)% at 34 degrees celsius; and (3.79 - or + 0.45)% at 32 degrees celsius; as compared to that in normothermia [(33.02 - or + 3.58)%] in the absence of rat white blood cells (WBC) (P<0.001). LDH release rate of the hypoxic cells further decreased in the presence of rat WBC to (51.14 - or + 2.17 )% at 37 degrees celsius;, (19.53 - or + 4.37)% at 34 degrees celsius; and (16.68 - or + 1.47)% at 32 degrees celsius; (P<0.001). In the scratched cells, with or without WBC, LDH release rate showed no significant variation between the 3 temperatures (P>0.05)., Conclusion: Mild hypothermia offers obvious protective effects on rat astrocytes against ischemic damage but not against mechanical injury.

Micro-Tom is a miniature dwarf tomato, which can grow at a high density, has a short life cycle, and can be transformed efficiently. As a result, it became a new model plant for functional genomics study. The origin and biological characteristics of Micro-Tom were summarized. Recent advances in the mechanisms involved in the miniature dwarf trait, as well as the application of Micro-Tom in plant functional genomics study and the improved genetic transformation systems were reviewed.

Objective: To study the therapeutic effects of koumine on psoriasis in mouse models., Methods: The effects of koumine on epithelial cell mitosis and epidermal cell differentiation was evaluated by collecting the samples of the vaginal mucous and squamous epidermis at the tail of mice treated with methotrexate or koumine at different doses. The levels of interleukin (IL)-2 were detected with enzyme-linked immunosorbent assay., Results: High and intermediate doses of koumine showed remarkable inhibitory effect on mouse vaginal epithelial cell mitosis and promoted the formation of epidermal granular layer in the scales at the mouse tail. Three concentrations at 6, 30, 150 mg/kg of koumine decreased serum IL-2 level in the mice., Conclusion: The therapeutic effect of koumine against psoriasis is related to the inhibition of epidermal cell proliferation, promoting the formation of granular cells and decreasing the serum level of IL-2.

Objective: To assess the separation efficiency of magnetic-activated cell sorting in the purification of CD4+ T cells from murine spleen, and observe the effects of koumine on the proliferation of the separated cells., Methods: CD4+ T cells were isolated from murine spleen by magnetic-activated cell sorting (MiniMACS). Fluorescence-activated cell sortering was employed to determine the purity of CD4+ T cells before and after the separation procedure followed by evaluation of the cell viability using trypan blue staining. Concanavalin A- (ConA, 5 microg/ml) or phytahematoagglutinin (PHA,1 mg/ml)-induced murine T cells were treated with different concentrations of koumine (10-320 microg/ml), and their proliferation was determined by MTT colorimetry, and enzyme-linked immunosorbent assay was used to measure IL-2 level in the cell culture supernatant., Results: The purity of CD4+ T cells reached (90.3+/-5.8)% after the purification with a cell viability of (94.9+/-3.6)%. Koumine (20-320 microg/ml) dose-dependently inhibited ConA- or PHA-induced proliferation of murine lymphocytes as compared with the controls (P<0.05). Koumine (20, 100, and 200 microg/ml) significantly decreased the level of IL-2 in comparison with the control group (P<0.05)., Conclusions: CD4+ T cells of high purity can be obtained from murine spleen using MiniMACS without impairing the viability of the cells. Koumine significantly inhibits the proliferation of murine CD4+ T cells due to its immunosuppressive effect and inhibition of IL-2 secretion.

Objective: To identify the structure of the alkaloid extract of Gelsemium from east Guangdong province of China., Methods: Fourier transform infrared (FTIR) absorption spectrometry, hydrogen and carbon spectra of nuclear magnetic resonance (NMR) and distortionless enhancement by polarization transfer (DEPT) analysis were used for the structural identification of the alkaloid exstract of Gelsemium., Results: The frequency, intensity and shape of the extract's characteristic peaks in infrared absorption spectra (4,000.0-400.0 cm(-1)) and (1)H NMR, (13)C NMR were recognized and compared. The molecular structure of the sample was consistent with the theoretically derived model., Conclusion: The extract is structurally identical to koumine, which may provide evidence for its safe clinical application and establishment of Chinese medicine fingerprint database of Gelsemium.

Objective: To establish a method for isolating and identifying koumine., Methods: Koumine was first extracted from Gelsemium alkaloids by using chloroform, which was further separated and purified by column chromatograph, and identified by thin-layer chromatography., Results and Conclusion: Koumine crystals were isolated and purified from Gelsemium alkaloids effectively.

Objective: To develop a simple and efficient method for extracting Gelsemium alkaloids., Methods: Uniform design and orthogonal design were respectively utilized to optimize the 3 factors in the extraction process, namely the type of solvent used, the solid-to-liquid ratio in the extraction system and the reflux time., Results: Both of the design methods adopted trichloromethane as the solvent, with the solid-to-liquid ratio of 1:7.5 and reflux time of 3 h with 3 repetitions for the highest yield of alkaloids., Conclusion: Similar results are obtained from the two design methods for extracting Gelsemium alkaloids.