In order to investigate the comprehensive quality differences of the seeds harvested in different growing time, and offer theoretical guide for the optimization of seeds' production technology, we analyzed the apparent size, 1 000-grain weight, water absorbency, germination indexes, postharvest embryo rate change, relatively contents of coumarins and the yield of single plant of its seeds of different harvesting time, and compared their comprehensive quality by Topsis analysis. The results showed that there existed obvious differences in yield and quality between seeds of 3 harvesting times. While the harvesting time postponed, the yield of single plant raised, and the shrunken seeds rate decreased, the quality of seedlings increased, while the contents of coumarins showed a steady increase, and the germination rate decreased. The comprehensive quality of the seeds harvested in the black ripe time rank the first place, followed by the brown ripe time and the yellow ripe time. As the harvesting time delays, the seeds' comprehensive quality increases, therefore, we could put off the seeds' harvesting time properly for the high efficient seed production.
Bupleurum ; growth & development ; metabolism ; physiology ; Germination ; Plant Proteins ; chemistry ; metabolism ; Quality Control ; Seeds ; growth & development ; metabolism ; physiology ; Solubility ; Time Factors ; Water ; metabolism

OBJECTIVE: To explore the effect of delta-opioid receptor (DOR) in electroacupuncture (EA) protecting the brain against acute ischemic injury. METHODS: Fifty-one rats were randomly divided into sham ischemia group, ischemia group, sham EA group, EA group, and EA+DOR antagonist (naltrindole) group. Transient focal cerebral ischemia (1 hour) was induced in rat brain by middle cerebral artery occlusion (MCAO) method. EA was applied on Shuigou (GV 26) and Neiguan (PC 6) for 30 min, starting immediately after the onset of reperfusion. Neurological deficit scores and volume of cerebral infarction were detected after 24-hour reperfusion. Other 12 rats were randomly divided into sham ischemia group, ischemia group, EA group and EA + naltrindole group. DOR protein expressions were assessed by Western blotting after 24-hour reperfusion. RESULTS: In comparison with the ischemia group and sham EA group, EA significantly reduced ischemic infarction and neurological deficits (P<0.05); EA significantly increased the expression of 60 kD DOR protein (P<0.05) and tended to increase that of 36 kD DOR protein (P>0.05). When naltrindole was combined with EA, the naltrindole completely abolished the EA-induced protection in ischemic infarction and neurological deficits, and also arrested the expression of DOR. CONCLUSION: EA can up-regulate DOR expression and protect the brain from ischemia-reperfusion injury.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

Background:A ruptured hepatocellular carcinoma (HCC) is often fatal.In addition to surgery and transarterial embc lization,radiofrequency ablation (RFA) might be another option for treating a ruptured HCC.Unfortunately,conventional RFA has a limited ablation zone;as such,it is rarely used to treat ruptured tumors.Case presentation:This case was a 60-year-old man who had a large,ruptured HCC in which hydrochloric acid (HCI)-enhanced RFA successfully controlled the bleeding and made the tumor completely necrotic.Conclusion:Considering the effectiveness of HCI-enhanced RFA in achieving hemostasis and tumor ablation,it might be a new option for treating large,ruptured HCCs.

OBJECTIVETo study the expression changes of tumor necrosis factor-alpha (TNF-(alpha), interleukin-6 (IL-6), C-reactive protein (CRP), monocyte chemotactic protein 1 (MCP-1), and their correlation with obesity in 20 -50 years old population of phlegm-damp constitution (PDC) and of normal constitution (NC) using Luminex technique.

METHODSTotally 101 population were recruited from Health Examination Center of Puren Hospital from April to December 2011. Based on body mass index (BMI), the subjects were assigned to four groups, i.e., the obesity of PDC group (Group OBT, BMI > or = 24 kg/m2, 30 cases), the non-obesity of PDC group (Group NOBT, BMI < 24 kg/m2, 25 cases), the obesity of non-PDC group (Group OBNT, BMI > or = 24 kg/m2, 28 cases), the NC group (Group P, BMI < 24 kg/m2, 18 cases). The BMI and body fat percent (FAT%) were compared among the 4 groups. Serum levels of TNF-alpha, IL-6, CRP, and MCP-1 were measured with Luminex technique.

RESULTSBMI was significantly higher in Group OBT and Group OBNT than in Group NOBT and Group P (all P < 0.05). The FAT% was significantly higher in Group OBT and Group OBNT than in Group P (P < 0.01). The serum TNF-alpha level in Group OBT was higher than in Group P (P < 0.01). The serum CRP and MCP-1 levels were significantly higher in Group OBT, NOBT, and OBNT than in Group P (P < 0.05, P < 0.01). The score for PDC was positively correlated with TNF-alpha, IL-6, and MCP-1 levels (P < 0.05).

CONCLUSIONSAbnormal higher levels of inflammatory factors exist in 20 -50 years old population of PDC. Chronic inflammation exists in population of PDC and obesity people.

Adult ; Body Mass Index ; C-Reactive Protein ; metabolism ; Case-Control Studies ; Chemokine CCL2 ; blood ; Female ; Humans ; Inflammation ; Interleukin-6 ; blood ; Male ; Medicine, Chinese Traditional ; Microchip Analytical Procedures ; Middle Aged ; Obesity ; blood ; diagnosis ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

microRNA is specifically expressed in ocular tissues and plays a unique role in ocular tissues. microRNA- 155 regulates a variety of signaling pathways, affects the expression of the corresponding protein,then leads to the morphology and function abnormality of cells and tissue. The mechanism of primary open angle glaucoma is not yet clear. It is considered that the morphology and function abnormality of trabecular meshwork is the main cause of the increasing intraocular pressure. The effect of microRNA- 155 on primary open angle glaucoma can provide a new direction for the pathogenesis, diagnosis and treatment of primary open angle glaucoma.

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor,recombinant CHO cell strain MK3-A2 was cultured,which could secrete rhTNK-tPA, by a serum-free perfusion culture system. The culture periods in this two experiments were as long as 77 and 110 days respectively. The cells density reached 2?107 cells /ml. The average volumetric productivity of rhTNK-tPA was 89 mg/L?d, and the highest one was 216mg/L?d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570?m. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.

Objectlve To detect the KChIP2 mRNA level in rheumatic heart disease patients with or without atrial fibrillation (AF) by real-time PCR.Methods Right atrial appendage samples from rheumatic heart disease patients with (n=17) or without AF (n=13) were obtained during cardiac surgery.Total RNA was extracted from the atrial tissues.and the KChIP2 and Kv4.3 mRNA were detected by SYBR Green I real-time PCR with the GAPDH as the house keeping gene.Result The ratio of KChIP2/GAPDH(0.1468 ±0.0452 VS.0.2200±0.0388,P＜0.01)and the ratio of Kv4.3/GAPDH(0.3946±0.1826 vs.0.5257±0.1427.P＜0.05)were significantly lower in AF patients compared to non-AF patients.Conclusion Down-regulated atrial KChIP2 and Kv4.3 mRNA expressions in rheumatic heart disease patients with chronic AF might be one of the molecular bases responsible for the down-regulation of the Ito current density of AF.

Objective To establish the fingerprint of Fangshu Qingre mixture and evaluate its quality in combination with chemical pattern recognition.Methods Using 3,5-O-dicaffeoyl quinic acid as the reference peak,ultra performance liquid chrmatography(UPLC)fingerprint of 10 batches of Fangshu Qingre mixture were established with Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition).The common peaks were identified and similarity evaluation were conducted,to determine the attribution of each common peak.The cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least squares method-discriminant analysis(OPLS-DA)in the chemical pattern recognition method were used to determine the differential components affecting the quality of the preparations.Results There were 31 common peaks in the fingerprints of 10 batches of Fangshu Qingre mixture,and their similarities were between 0.975 and 0.996.Six common peaks were identified,including peak 9(chlorogenic acid),peak 19(luteolin-7-O-β-D-glucoside),peak 21(3,5-O-dicafeoyl quinic acid),peak 23(hesperidin),peak 29(linarin),and peak 31(patchoulone).CA and PCA divided 10 batches of Fangshu Qingre mixture into 3 categories.The results of PCA showed that the cumulative variance contribution rate of principal components 1～6 was 95.947％.The results of OPLS-DA showed that there were 13 peaks VIP greater than 1,which was the difference component.Conclusion The method is simple and sensitive,can be used to evaluate the quality of Fangshu Qingre mixture.

OBJECTIVETo construct glypican-3 (GPC3)-green fluorescent protein eukaryotic expression vector pEGFP-c3-GPC3, and analyze the effect of GPC3 on the proliferation of human hepatoma cell line MHCC-97L.

METHODSThe eukaryotic expression vector pEGFP-c3-GPC3 was constructed with recombinant DNA technique and transfected into MHCC-97L cells via Lipofectamine 2000. The cells stably expressing GPC3 were screened by flow cytometry and G418. The mRNA expression of GPC3 was detected by RT-QPCR method, and the protein expression by Western blotting and fluorescence microscope. The effect of GPC3 gene on the growth of the cells was examined by MTT assay.

RESULTSRestriction endonuclease analysis and DNA sequencing verified correct construction of the recombinant plasmid. The green fluorescence was detected in the transfected MHCC-97L cells under fluorescence microscope. RT-QPCR and Western blotting both confirmed successful expression of GPC3 in MHCC-97L cells. The growth curve showed a significant acceleration of the proliferation of the transfected MHCC97-Lsol;GPC3 cells as compared with MHCC97-L and MHCC97-L/C3 cells (P<0.001).

CONCLUSIONWe have successfully constructed the eukaryotic expression vector pEGFR-c3-GPC3, which allows stable GPC3 expression in MHCC97-L/GPC3 cells. The upregulation of GPC3 expression can stimulate the growth of hepatoma cell line MHCC97-L in vitro.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Genetic Vectors ; Glypicans ; pharmacology ; Green Fluorescent Proteins ; genetics ; Humans ; Liver Neoplasms ; pathology ; Plasmids ; Transfection