OBJECTIVETo investigate the apoptosis and proliferation effect of matrine on human medulloblastoma cell line D341 in vitro and the effect of the expression of the related caspase 3 and caspase 9 proteins.

METHODSThe D341 cells were cultivated successfully in vitro. Then the cells were divided into 5 groups according to the concentration of matrine (0.5 mg/mI group, 1.0 mg/ml group, 1.5 mg/ml group, 2.0 mg/ml group and the control group was 0 mg/ml). All the experiments were repeated three times. The cell morphologic and structure change was observed with the optical microscope and the transmission electron microscope. The proliferation of D341 cell was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining. The expression of Caspase3 and Caspase9 was detected by Western blot.

RESULTSWith the effect of matrine, the proliferation inhibition rate gradually increased with drug concentrations increasing, and there was a significant difference (P < 0.01). The inhibitory effect of matrine on cell proliferation was different with the different treatment time, there was a significant difference between the 24 h to 72 h groups (P < 0.01). The apoptotic rate increased with matrine concentrations increasing. There were significant differences between the group of 0.5 mg/mI or 1.0 mg/mI to the group of 1.5 mg/mI or 2.0 mg/mI (P < 0.05). The apoptotic rate increased with the prolonged treatment time. There were significant differences between the group of 24 h or 48 h to the group of 72 h ( P < 0.05). With the increase of matrine concentration, the expression of Caspase 3 and Caspase 9 increased (P < 0.01).

CONCLUSIONMatrine induces the apoptosis, and inhibits the proliferation of human medulloblastoma D341 cells in vitro by up-regulation of the expression level of Caspase3, Caspase9.

Alkaloids ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; metabolism ; pathology ; Humans ; Medulloblastoma ; metabolism ; pathology ; Quinolizines ; pharmacology ; Up-Regulation

Objective To investigate current status and problems of coagulation factor Ⅷ and Ⅸ assay in domestic laboratories so as to provide the reference for implementing the standardization and quality improvement.Methods A questionnaire survey was carried out in 76 laboratories,and quality control materials were distributed to 54 laboratories for activity assay.The questionnaire information was analyzed statistically.Test results of quality control materials were classified into three groups according to the reagents and the ranked grading analysis were used to evaluate the performance.Results This research was investigative study.The amount of sample was less than 30 per month in 72％ (52/72)of laboratories.The frequencies of calibration were different,and 33％ (24/72) of laboratories did not perform calibration in a different assay batch.39％ (28/72)of laboratories did not run internal quality control,and about 21％ (15/ 72) of laboratories just performed the normal level quality control.Individual laboratories showed a high cumulative CV (＞ 30％) of intemal quality control.For normal FⅧ and FⅨ control materials,the CV of results were 11.3％-18.2％ and 11.3％-17.9％ respectively as well as 15.3％-20.3％ and 19.5％-21％ for abnormal.Of the three groups,the proportions of laboratories which the FⅧ test results out with consensus were18％,24％ and 22％ as well as 20％,24％ and 28％ for FⅨ.Conclusions The key requirements for quality control of coagulation factors active assay remain to be addressed and implemented.The repeatability and comparability in some laboratories are not satisfactory to meet the clinical needs.With the purpose of promoting quality improvement,we need to develop guidelines,organize related training and establish a national external quality assessment scheme.

Objective To explore clinical features and antimicrobial resistance of Klebsiella pneumoniae (K.pneumoniae) lower respiratory tract infection(LRTI) in children.Methods Clinical data of 107 children with K.pneumoniae LRTI confirmed by sputum culture from January to December 2015 were analyzed retrospectively.Results 62.62% of children with LRTI were aged less than 6 months and 64.49% episodes occurred in autumn and winter.All cases had cough and 39 had fever, the main complications were type I respiratory failure, type Ⅱ respiratory failure, cardiac insufficiency, and electrolyte disturbance, 39 cases(36.45%) had complications involving two systems, 5 cases(4.67%)had complications involving three systems,47 cases (43.93%) met the diagnostic criteria of severe pneumonia.43 cases (40.19%) had primary underlying diseases, the major were congenital heart disease, preterm and low birth weight, and malnutrition.Children with imipenem-resistant bacteria infection were more prone to develop extrapulmonary complications than those with non-resistant pathogenic infection.The resistance rate of K.pneumoniae to amikacin was the lowest(9.35%).90 cases were recovered and markedly effective, 11 cases were effective, 4 cases were not healed and voluntarily discharged from hospital, and 2 cases died.Conclusion Children aged less than 6 months and with underlying diseases are prone to develop LRTI, and complications are more.

Objective: To determine the changes of serum Tau protein, glial fibrillary acidic protein (GFAP), tumor necrosis factor alpha (TNF-α), and malonaldehyde (MDA) in rats after blast-related traumatic brain injury (BTBI) and to provide relative information for further studies on BTBI mechanism and seek specific biomarkers for BTBI.
Methods: Ninety male Sprague-Dawley rats were randomly assigned into three groups: control group, moderate blast injury group, and severe blast injury group (n=30 for each). Rats in the moderate and severe blast injury groups were respectively exposed to corresponding levels of BTBI. After explosion, serum levels of Tau, GFAP, TNF-α, and MDA in each group were determined by Elisa assay at different time points after injury (8 h, 24 h, 3 d, and 6 d). The extent of brain damage was detected by Nissl staining and TUNEL assay.
Results: Serum levels of Tau and GFAP rapidly increased and reached the peak at 24 h after either moderate or severe blast injury. All the values were significantly higher than control group at all time points (P<0.05). Serum TNF-α level of both injury groups peaked at 8 h after BTBI and stayed significantly higher than control group at all time points (P<0.05). Serum MDA of two injury groups began to significantly increase at 3 d and the level stayed significantly higher than control group until 6 d (P<0.05). Moreover, unlike the other biomarkers, serum MDA of severe blast injury group was significantly higher than moderate blast injury group at 6 d (P<0.05).
Conclusion: The changes of serum Tau, GFAP, and TNF-αshowed a good sensitivity at the acute phase after BTBI (within 24 h). However, their specificity and correlation with the extent of injury were limited in this experiment. Moreover, although the change of serum MDA showed a poor sensitivity and specificity to the diagnosis of BTBI during the first few days, it can reflect the injury degree at 6 d after injury. Therefore, further studies are needed to improve the methods of detecting more serum markers and investigate the significance of multiple markers in diagnosing BTBI.

Objective To extract the molluscicidal active component, Eomecon Chionantha Hance Alkaloids (ECA)from Eomecon Chionantha Hance. Methods The molluscicidal active component from the rootstalk of Eomecon Chionantha Hance was extracted and purified by using the method, 70% alcohol percolation, decompress concentrating, salting-out and so on. Results The final product rate of ECA was 1.009%. Conclusion This method is economic, simple and reliable, and suitable to industrialized production.

Objective To explore the expression of PIWIL1 protein and DICER enzyme in hepa tocellular carcinoma (HCC) and their significance.Methods Immunohistochemical method was used to detect the expression of PIWIL1 and DICER in 47 cases of HCC and the adjacent HCC tissues.Western blot method was used to detect the expression of PIWIL1 and DICER in 31 cases of fresh HCC tissues and their adjacent HCC tissues.The relationship between PIWIL1 and DICER and their relationships were analysed with clinical features.12 cases of normal liver tissues were used as control group.Results The expression of PIWIL1 was high in HCC but low in normal liver tissues (P＜ 0.05).The expression of DICER was high in normal liver tissues but low in HCC (P＜0.05).The expression of PIWIL1 was positively correlated with invasion to adjacent tissues and histological differentiation (P＜0.05).The expression of DICER was negatively correlated with invasion to the adjacent tissues and histological differentiation (P＜0.05).There was a negative correlation between PIWIL1 and DICER (P＜ 0.05).Conclusions High expression of PIWIL1 and low/missing expression of DICER was related to pathological differentiation and invasion of adjacent tissues.

Objective To study the distribution of Oncomelania hupensis snails in mountainous regions and the dynamic charaeteristics of the distribution.Methods An environment calledLanbaoclosed to Puge County.Sichuan Provinee WaS selected as the study field.Random sampling was designed to determine the investigation sites.The snails were collected and the hying snaila were identified by the method of dissection in the laboratory.The distribution of snails was analyzed by some statistical indices,such as mean,variance and so on.Then the negative binomial distribution.log-normal distribution and exponential distribution were fitted to the snail data by the method of maximum likelihood estimation to explore the snail distribution in different time.Results The negative binomial distribution was fitted well to the snail data in April,May,July,August,September,November in 2008,and no distribution was fined to the snail data in June,October.December in 2008 and February in 2009.Conclusions The distribution of Oncomelania hupensis in mountainous regions is not simple negative binomial distribution,but pwbably a dynamic process and an uncertain distribution.

Exploring the influence of extract of Ginkgo biloba (EGB) on the proliferation, apoptosis of ACC-2 cell in lacrimal adenoid cystic carcinoma and analyzing the influence of EGB on the gene expression of Survivin and TIP30 based on the levels of the gene and protein. ACC-2 cell in human with ACC of lacrimal gland disposed by EGB of different concentration was in vitro cultured. MTT method was used for cell proliferation detection. Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle. Survivin and TIP30 gene expression together with protein expression were analyzed by RT-PCR and Western blotting. And it is indicated that EGB has inhibitory effect on the proliferation of ACC-2 cell in vitro. Furthermore, the dose-effect relationship was significant. Compared with the control group, it had statistical difference (P <0.01). The inhibitory concentration 50% (ICso) is 88 mg . L-1. By flow cytometer examination, it was indicated that EGB can gradually increase ACC-2 cell in G0-G1 stage and decrease it in G2-M and S stage. With the increase of dose, the apoptosis rate of ACC-2 cell obviously increased (P <0.05 or P <0.01). Both of the expression results of RT-PCR and Western hybrid proteins have showed that the concentration of EGB increased, it could be seen a significant decrease in Survivin gene expression (P <0.01). Meanwhile, the TIP30 gene expression got a significant increase. Therefore, EGB can effectively inhibit ACC-2 cell Survivin gene expression in human with adenoid cysistic carcinoma of larcrimal gland as well as promoting TIP30 gene expression, inducing the ACC-2 cell apoptosis and inhibiting tumor cell proliferation, which provided a certain theoretical and experimental basis for the application of Chinese herbal medicinal ingredient in the treatment of tumors.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Carcinoma, Adenoid Cystic ; drug therapy ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; Gene Expression ; Ginkgo biloba ; chemistry ; Humans ; Lacrimal Apparatus ; drug effects ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology

AIM:To evaluate the effects of treatment with HP 1188-immunoglobulin yolk ( HP1188-IgY) on Helicobacter pylori ( H.pylori)-infected gastritis in BALB/c mice.METHODS:BALB/c mice were used to establish an animal model of H.pylori-infected gastritis, and the mice were divided into 8 groups (10 mice per group).Oral antibiotics were used in group 1, 1 mg HP1188-IgY in group 2, 1 mg HP1188-IgY plus 30%sucralfate in group 3, 5 mg HP1188-IgY in group 4, 5 mg HP1188-IgY plus 30%sucralfate in group 5, PBS in group 6, and 30% sucralfate in group 7 with the treatment once per day for 10 d;and 2.5 mg HP1188-IgY was injected hypodermically twice with a 48-h interval in group 8.Another 10 mice were used as normal control in group 9.The planting of bacteria in the stomach was assayed by bacteri-al culture, rapid urease test, PCR and pathological sectioning .RESULTS:Intragastric administration with 1 mg HP1188-IgY plus 30%sucralfate per day effectively cured the injury of gastric mucosa caused by H.pylori infection, and the effect has no significant difference compared with antibiotics (P>0.05).CONCLUSION:We establish a BALB/c mouse mod-el infected with H.pylori successfully.Sucralfate (30%) is an ideal protectant for HP1188-IgY, which might decrease H. pylori infection in the stomach of BALB/c mice by oral inoculation .

Objective Cordycepin exhibits varieties of pharmacological effects including anti-pathogen, anti-inflammatory, immune-modulatory, and anti-cancer activities.Recent studies indicate cordycepin may also have significant therapeutic potential in many diseases, such as metabolic disorders, oxidative injury and central nervous system diseases through different mechanisms , which are gradually clarified.The current progress and future prospects are reviewed in this paper.