Production of all-male non-transgenic zebrafish by conditional primordial germ cell ablation.

Many fish species exhibit remarkable sexual dimorphism, with males possessing numerous advantageous traits for commercial production by aquaculture such as faster growth rate, more efficient food energy utilization for muscle development, and better breeding performance. Several studies have shown that a decrease in the number of primordial germ cells (PGCs) during early development leads predominantly to male progeny. In this study, we developed a method to obtain all-male zebrafish (Danio rerio) by targeted PGC ablation using the nitroreductase/metronidazole (NTR/Mtz) system. Embryos generated by female heterozygous Tg(nanos3:nfsB-mCherry-nanos3 3'UTR) and male wild-types (WTs) were treated with vehicle or Mtz. Compared to vehicle-treated controls, 5.0 and 10.0 mM Mtz treatment for 24 h significantly reduced the number of PGCs and yielded an exclusively male phenotype in adulthood. The gonads of offspring treated with 5.0 mM Mtz exhibited relatively normal morphology and histological characteristics. Furthermore, these males were able to chase females, spawn, and produce viable offspring, while about 20.0% of males treated with 10.0 mM Mtz were unable to produce viable offspring. The 5.0 mM Mtz treatment protocol may thus be suitable for large-scale production of fertile male offspring. Moreover, about half of these males were WT as evidenced by the absence of nfsB gene expression. It may thus be possible to breed an all-male WT fish population by Mtz-mediated PGC ablation.

ZNF655 mediated by LINC01210/miR-124-3p axis promotes the progression of gastric cancer.

Gastric cancer (GC) is a common malignant tumor that usually originates from the epithelium of the gastric mucosa. ZNF655 was a suppressor gene of many cancers. However, the mechanism of ZNF655 in GC remains unknown. Quantitative polymerase chain reaction was used to assess the expression of ZNF655, LINC01210, and miR-124-3p. Western blotting was used to monitor ZNF655 protein expression. MTT, clone formation, transwell, and flow cytometry were all used to investigate the functions of GC cells. The interactions between ZNF655, LINC01210, and miR-124-3p were confirmed using the dual-luciferase reporter gene assay and the RIP assay. ZNF655 was highly expressed in GC cells. ZNF655 knockdown reduced GC cell viability, proliferation, migration, invasion, and induced apoptosis. The level of miR-124-3p was significantly reduced in GC cells. Besides, miR-124-3p targeted ZNF655 and inhibited its expression. MiR-124-3p mimics inhibited GC cell progression, but ZNF655 overexpression reversed these effects. Moreover, LINC01210 was found to be highly expressed in GC cells and to be able to sponge miR-124-3p. Furthermore, inhibiting miR-124-3p or increasing ZNF655 could counteract the effects of LINC01210 knockdown on GC cell development. Finally, ZNF655 promoted GC cell progression and was regulated by the LINC01210/miR-124-3p axis.

Physicochemical properties and bioactivities of original and Se-enriched polysaccharides with different molecular weights extracted from Pleurotus ostreatus.

Three polysaccharides (WZP1, WZP2, WZP3) and their Se-enriched products (SeWZP1, SeWZP2 and SeWZP3) were obtained from Pleurotus ostreatus using a simple, rapid method and HNO3-Na2SeO3 method, respectively. The molecular weight distribution profiles of all samples except SeWZP2 showed double peaks. The average molecular weights (Mw) of WZP1-3 were 48.6 kDa, 20.2 kDa and 11.8 kDa, respectively, and of SeWZP1-3 were 19.6 kDa, 37.7 kDa, 14.5 kDa, respectively. The complexity of monosaccharide composition of WZP1-3 was inversely proportional to the ethanol concentration used in the ethanol precipitation process. Additionally, the results of biological activity tests indicated that α-glucosidase inhibitory activity of WZP1-3 was related to the molecular weight and the monosaccharide composition complexity. The selenized modification can improve the α-glucosidase-inhibiting, hydroxyl radical-scavenging activity of P. ostreatus polysaccharides. Therefore, by improving their bioactivities by selenization, the polysaccharides of P. ostreatus could be utilized as a natural health food supplement.

Research on rat's pulmonary acute injury induced by lunar soil simulant.

BACKGROUND: The steps to the moon never stopped after the Apollo Project. Lessons from manned landings on the moon have shown that lunar dust has great influence on the health of astronauts. In this paper, comparative studies between the lunar soil simulant (LSS) and PM2.5 were performed to discover their harm to human biological systems and explore the methods of prevention and treatment of dust poisoning for future lunar manned landings. METHODS: Rats were randomly divided into the control group, two CAS-1 lunar soil simulant groups (tracheal perfusion with 7 mg and 0.7 mg, respectively, in a 1-mL volume) and the PM2.5 group (tracheal perfusion with 0.7 mg in a 1-mL volume). The biochemical indicators in the bronchoalveolar lavage fluid (BALF), MPO activity in the lung tissue, pathologic changes, and inflammatory cells in the BALF were measured after 4 h and 24 h. RESULTS: The LSS group showed cytotoxicity that was closely related to the concentration. The figures of the two LSS groups (4 and 24 h) show that the alveolar septa were thickened. Additionally, it was observed that neutrophils had infiltrated, and various levels of inflammation occurred around the vascular and bronchial structures. CONCLUSION: The overall results of the acute effects of the lungs caused by dust showed that the lung toxicity of LSS was greater than that of PM2.5. LSS could induce lung damage and inflammatory lesions. The biomarkers in BALF caused by acute injury were consistent with histopathologic observations.

[Influence Factors and Sensitivity of Ozone Formation in Langfang in the Summer].

The diurnal variations in ozone concentrations in the summer were studied using the temperature, wind speed and direction, total cloud cover, and solar radiation intensity data collected in Langfang, China. The ratio of volatile organic compounds to nitrogen oxides (VOCs/NOx) and the EKMA curve were studied to analyze the sensitivity of ozone formation. The results showed that:① The ozone generation rate and ozone concentrations were positively correlated with the solar radiation intensity with Pearson correlation coefficients of 0.61 and 0.48, respectively. Both the ozone generation rate and the solar radiation intensity reached their peak at about 12:00, while the ozone concentration reached its peak at about 16:00, which lagged behind the peak of the solar radiation intensity by 4 h. ② The correlation coefficient between the ozone generation rate and the temperature was 0.44, between the ozone concentration and temperature was 0.68. The ozone generation rate and ozone concentrations were inversely correlated with total cloud cover with correlation coefficients of -0.24 and -0.45, respectively. ③ The ozone concentrations in Langfang were high when the winds were from the west, south, or southeast. ④ The ozone concentrations in Langfang were more sensitive to VOCs than to NOx; thus, they can be reduced efficiently by controlling the VOCs emissions.

The Dynamics and Regulatory Mechanism of Pronuclear H3k9me2 Asymmetry in Mouse Zygotes.

H3K9 methylation is an important histone modification that is correlated with gene transcription repression. The asymmetric H3K9 dimethylation (H3K9me2) pattern between paternal and maternal genomes is generated soon after fertilization. In the present study, we carefully determined the dynamics of H3K9me2 changes in mouse zygotes, and investigated the regulatory mechanisms. The results indicated that histone methyltransferase G9a, but not GLP, was involved in the regulation of asymmetric H3K9me2, and G9a was the methyltransferase that induced the appearance of H3K9me2 in the male pronucleus of the zygote treated with cycloheximide. We found that there were two distinct mechanisms that regulate H3K9me2 in the male pronucleus. Before 8 h of in vitro fertilization (IVF), a mechanism exists that inhibits the association of G9a with the H3K9 sites. After 10 h of IVF the inhibition of G9a activity depends on yet unknown novel protein(s) synthesis. The two mechanisms of transfer take place between 8-10 h of IVF, and the novel protein failed to inhibit G9a activity in time, resulting in the appearance of a low level de novo H3K9me2 in the male pronucleus.

[Fragmentation pathways of five furocoumarine using line ion trap with orbitrapmass spectrometry].

OBJECTIVE: The multiple levels fragmentations of five furocoumarine (psoralen, xanthotoxin, bergapten, oxypeucedanin, and byakangelicol) in Angelica dahurica have been demonstrated using LTQ-Orbitrap mass spectrometry with high resolution and high mass accuracy to discover the possible,fragmentation regularity. METHOD: Duringcollsion-induced dissociation (CID), the MS(n) data of the five compoundswhich were gained in the positive ion mode at 35ev collision energy by direct injection syrings method were analyzed using Xcalibar 2.0 Software to infer the formula of these fragmentations. RESULT: The results indicated that the five compounds have similar fragmentation process with CO meutral lost at C5,C8-subsituents and furan ring, meanwhile the meutralloss of CO2 occurred easily at lactone group. CONCLUSION: This method is helpful in identifying the structures of other furocoumarinein Angelica dahuricaand their metabolites in vivo.

Qualitative and Quantitative Analysis of Lignan Constituents in Caulis Trachelospermi by HPLC-QTOF-MS and HPLC-UV.

A high-performance liquid chromatography coupled with quadrupole tandem time-of-flight mass (HPLC-QTOF-MS) and ultraviolet spectrometry (HPLC-UV) was established for simultaneous qualitative and quantitative analysis of the major chemical constituents in Caulis Trachelospermi, respectively. The analysis was performed on an Agilent Zorbax Eclipse Plus C18 column (4.6 mm×150 mm, 5 µm) using a binary gradient system of water and methanol, with ultraviolet absorption at 230 nm. Based on high-resolution ESI-MS/MS fragmentation behaviors of the reference standards, the characteristic cleavage patterns of lignano-9, 9'-lactones and lignano-8'-hydroxy-9, 9'-lactones were obtained. The results demonstrated that the characteristic fragmentation patterns are valuable for identifying and differentiating lignano-9,9'-lactones and lignano-8'-hydroxy-9,9'-lactones. As such, a total of 25 compounds in Caulis Trachelospermi were unambiguously or tentatively identified via comparisons with reference standards or literature. In addition, 14 dibenzylbutyrolatone lignans were simultaneously quantified in Caulis Trachelospermi by HPLC-UV method. The method is suitable for the qualitative and quantitative analyses of dibenzylbutyrolatone lignans in Caulis Trachelospermi.

[Textual research on the edition of WU Kun's books].

WU Kun is a famous physician of the Xin'an School. He wrote a 6-volume Yi fang kao (Textual Research on Recipes) in 1584, a 24-volume Wu zhu su wen (Wu's Annotation of Plain Questions) in 1594, a 6-volume Zhen fang liu ji (Six Collections of Acupuncture and Prescription) in 1618 block-printed and funded by CHENG Biao, and a 2-volume Mai yu (Language of Pulse) with writing date unknown. For Yi fang kao, there were 4 editions, including Ming-dynasty block-printed edition, Japanese and Korean block-printed edition, old hand-copied edition and modern printing edition. For Wu zhu su wen, there were 5 editions, Ming-Qing-dynasty block-printed edition, Japanese block-printed edition, old hand-copied edition, photocopy and modern edition. For Zhen fang liu ji, there were Ming-dynasty block-printed edition, hand-copied edition, photocopy and modern edition. Mai yu always was printed together with Yi fang kao, and its editions were same as those of Yi fang kao, except some modern printing edition with Mai yu excluded.

TBP dynamics during mouse oocyte meiotic maturation and early embryo development.

To maintain cell lineage, cells develop a mechanism which can transmit the gene activity information to the daughter cells. In mitosis, TBP (TATA-binding protein), a transcription factor which belongs to TFIID was associated with M phase chromosomes and was proved to be a bookmark for cellular memory. Although previous work showed that TBP was dispensable for mouse oocyte maturation and early embryo development, exogenous TBP protein was detected in the nuclear of oocytes and early embryos. It is still unknown whether exogenous TBP can associate with condensed chromosomes during meiosis and mouse early embryo development. In present study by the injection of GFP-tagged TBP mRNA we for the first time investigated TBP dynamics in mouse early embryos and confirmed its localization pattern in oocytes. The exogenous TBP enriched at germinal vesicle at GV stage but disappeared from the chromosomes after GVBD. Moreover, exogenous TBP was still dispersed from the chromosomes of somatic donor nuclear in oocytes by nuclear transfer (NT), further proving that oocyte has some mechanism to remove TBP. During mouse embryo development, the exogenous TBP was removed from the chromosomes of M phase zygotes, but was found to express weakly at the M phase of 2-cell. Moreover, in the blastocyst TBP was also detected at the M phase chromosomes. Overexpression of TBP caused the failure of oocyte maturation and embryo development. Our results supported the idea that TBP might be a marker for transmitting cellular memory to daughter cells.

[Research on blockprinted versions of Yokucho collected at the libraries in mainland China].

There are several blockprinted versions of Yokucho written by Yoshimasu Todo, a representative of the Koho School, spread into inland China. By comparison, it is found that all these versions use the printing blocks of "Borotei's collected blocks". The descriptions for all these versions reveal as follows: Anhui Provincial Library: the 5(th) year of Yisi (1785) of Japanese Tenmei reign, a blockprinted edition of Langkha Bookstore, Yoshidashokento from the Borotei blocks; National Library: the 5(th) year of Yisi (1785) of Japanese Teimei reign, a blockprinted edition of Hei'an Bookstore (Royal Bookstore), Siwento Bookstore from the Borotei blocks; Library of China Academy of Chinese Medical Sciences: the 9(th) year of Renshen (1812) of Japanese Bunkha reign, a blockprinted edition of Kyoto Bookstore, Langkha Bookstore, Yoshidashoken Bookstore from the Borotei blocks; Library of Aademia Sinica: (1) the 9(th) year of Renshen (1812) of Japanese Bunkha reign, Khaga-wo Bookstore, Langkha Bookstore from the Borotei blocks; (2) the 5(th) year of Yisi (1785) of Japanese Tenmei reign, a blockprinted edition of Hei'an Bookstore (Royal Bookstore), Siwento Bookstore from the Borotei blocks; Library of Life Sciences, Shanghai Information Center for Life Sciences, Chinese Academy of Sciences: the 9(th) year of Renshen (1812) of Japanese Bunkha reign, a blockprinted edition of Kyoto Bookstore, Langkha Bookstore, Yoshidashoken Bookstore Borotei blocks.

[Textual research of Bencaogangmu Yizhilu].

Bencaogangmu Yizhilu was compiled by DAI Baoyuan according to Bencao Gangmu in the late Qing dynasty. Due to the limited circulation of the book, it was little studied by people in later ages. Errors could be found concerning the book's content, editions and the author's life. After in-depth research, we concluded that the author's name was Baoyuan (courtesy name Xintian and pseudonym Shouyu). He was born about in 1818 and the year of death was unclear. Compilation of the book was completed in 1885 and the book was published in 1887. Recording 2240 kinds of herbs, the book included 8 volumes, 7 of which were text and the 8(th) volume was the index. It has two editions-the Wu Yuansi added Shanfang printed edition (the 13th year of the Guangxu Period, 1887) and the copy edition of the Qing dynasty.

miR-26b promotes granulosa cell apoptosis by targeting ATM during follicular atresia in porcine ovary.

More than 99% of ovarian follicles undergo atresia in mammals, but the mechanism of follicular atresia remains to be elucidated. In this study, we explored microRNA (miRNA) regulation of follicular atresia in porcine ovary. A miRNA expression profile was constructed for healthy, early atretic, and progressively atretic follicles, and the differentially expressed miRNAs were selected and analyzed. We found that miR-26b, which was upregulated during follicular atresia, increased the number of DNA breaks and promoted granulosa cell apoptosis by targeting the ataxia telangiectasia mutated gene directly in vitro.

[Research on the interaction between GSTP1 polymorphism and Helicobacter pylori infection in patients with gastric intestinal metaplaisa].

OBJECTIVE: To detect the distribution of polymorphism of GSTP1 and infection rate of Helicobacter pylori (H. pylori) in different kinds of gastric intestinal metaplasia (IM) and to explore the relationship between entergenic factor-polymorphism of GSTP1 and extogenic factor-H. pylori infection, to determine the risk of subtype of IM. METHODS: There were 381 cases in total, including 143 CGS(+) and 238 IM. H. E. stain was used for pathological diagnosis. HID-AB-pH2.5 methods were used to classify the IM. ELISA method was used to detect the antibody of H. pylor-IgG. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used to analyze the genotype. RESULTS: Frequency of G genotype was higher in the IM group, when compared to the CGS(+) groups (P < 0.05). The positive rate of H. pylori was statistically higher than CGS(+) (P < 0.01). By effect-modified model analysis, GSTP1 gene polymorphsim and H. pylori infection presented a positive interaction in the stage IM, with the OR value as 9.386 (95%CI: 3.736 - 23.580) after adjusted by age and gender. The synergy index was 2.078 and the attributable proportion of interaction was 46.36%. The positive rate of H. pylori were statistically highter than CGS(+) group in subtype IMI, subtype IMII and III (P < 0.01). The frequency of G genotype was higher in the IMII and III group, when compared with the IMI groups (P < 0.01). By effect-modified model analysis, in the stage of IMII and III, GSTP1 gene polymorphsim and H. pylori infection also presented a positive interaction, with OR value as 24.487 (95%CI: 7.731 - 77.735) after adjusted by age and gender, with its synergy index as 1.844, and attributable proportion of interaction as 43.89%. CONCLUSION: The infection of H. pylori and polymorphism GSTP1gene appeared to be both the external and internal factors, respectively. In the stage of IM, GSTP1 gene polymorphsim and H. pylori infection also presented a positive interaction, expecially in the IMII and III.

[Polymorphism of glutathione S-transferase P1 and correlation there of with carcinogenesis risk of gastric intestinal metaplasia].

OBJECTIVE: To investigate the distribution of polymorphism of glutathione S-transferase P1 (GSTP1) in different kinds gastric intestinal metaplasia (IM), and the carcinogenesis risk of different types of IM. METHODS: Peripheral blood samples were collected from 87 gastric cancer patients, 87 intestinal metaplasia patients, and 87 healthy persons as controls. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used to analyze the distribution of different alleles of GSTP1. Biopsy specimens of gastric mucous membrane were collected by gastroscopy. Histochemical staining for mucin was used to identify the kinds of IM. RESULTS: The G allele rates of the IM and gastric cancer groups were both significantly higher than that of the control group (chi2=6.921, P=0.009; chi2=28.787, P=0.000). The risk of IM of those with G allele was 1.944 times higher then that of the normal controls (95% CI: 1.177-3.209), and the risk of gastric cancer of those with G allele was 3.605 times higher (95% CI: 2.217-5.863). The G allele rate of the gastric cancer group was significantly higher than that of he IM group (chi2=7.687, P=0.006). The G allele rates of the type II and III IM were significantly higher than that of the normal controls (chi2=9.981, P=0.001; chi2=8.845, P=0.002). The risk of type II IM of the subjects with G allele was 2.747 times higher than that of the normal controls (95% CI: 1.475-5.114), and the risk of type III IM of the subjects with G allele was 3.451 times higher (95% CI: 1.556-7.657). The risks of type IIIM, type III IM, and gastric cancer of the subjects with allele G of GSTP1 gene were 2.905, 3.650, and 3.813 times higher than those of the normal controls respectively (95% CI: 1.341-6.293, 1.455-9.153 and 1.953-7.444 respectively). CONCLUSION: The individuals with G allele show an increased risk of developing IM and gastric cancer, especially type II and III IM. IM patients with G allele have the genetic characteristics similar to those of gastric cancer, so they should be regarded as high-risk individuals of gastric cancer and followed up regularly.

5-AZA-2'-deoxycytidine (5-AZA-CdR) leads to down-regulation of Dnmt1o and gene expression in preimplantation mouse embryos.

5-AZA-2'-deoxycytidine (5-AZA-CdR) is a demethylating, teratogenic agent and a mutagen, which causes defects in the developing mouse and rat after implantation. Our previous data indicated that 5-AZA-CdR (0.2 and 1.0 muM) inhibited the development of mouse preimplantation embryos. Pronuclear embryos exposed to 5-AZA-CdR at the pronuclear stage were unable to form 8-cell embryos, while 2-cell-stage embryos exposed to 5-AZA-CdR only developed into uncompacted 8-cell-stage embryos. And there was no formation of blastocysts when 4-cell embryos cultured in 5-AZA-CdR. In our present study, we detected Dnmt1o protein and some developmental gene expression in order to find the reasons for the developmental arrest. Dnmt1o could not traffic to 8-cell nuclei as control when embryos were exposed to 5-AZA-CdR. Dnmt1o was in cytoplasm at 2-cell and 4-cell stages before and after treated with 5-AZA-CdR. Gene expression changes were also detected in this research. Our data indicated that connexin 31 (Cx31), connexin 43 (Cx43), connexin 45 (Cx45), E-cadherin (Cdh1) and beta-catenin (Ctnnb1) were all downregulated by 5-AZA-CdR. Cx31, Cx43 and Cx45 are members of connexins family, which have a central role in gap junctions. Cdh1 and Ctnnb1 are necessary for the foundation of tight junctions. Therefore, developmental arrest induced by 5-AZA-CdR may be caused by the failure of Dnmt1o cytoplasmic-nuclear traffic and the down-regulation of developmental gene expression. Normal compaction and blastocoel cavitation need Dnmt1o traffic to 8-cell nuclei and the right gene expression, especially the correlative genes in gap junctions and tight junctions.

Factors affecting the serum gastrin 17 level: an evidence-based analysis of 3906 serum samples among Chinese.

OBJECTIVE: To investigate the influence of gender, age, site of lesion, disease type and Helicobacter pylori (H. pylori) infection on the human serum gastrin-17 level and to study the diagnostic value of serum gastrin-17 in gastric precancerous lesions and gastric cancer. METHODS: Serum gastrin-17 and serum H. pylori IgG antibody were detected by the ELISA method. The different gastric disease groups were confirmed by endoscopy and histopathology. RESULTS: Among the 3906 serum samples according to the gender, age, site of lesion and the data of different gastric disease groups, the serum gastrin-17 level was markedly higher in people>or=60 years old than that in younger age groups. The serum gastrin-17 level increased progressively in the following order: healthy control group, nonatrophic gastritis group, gastric ulcer group, and the serum gastrin-17 level was higher in the atrophic gastritis with dysplasia group than that without it, the lowest level being in the gastric cancer group. Among the 2946 serum samples matched with the site of the lesion, the serum gastrin-17 level was higher in those with antral diseases than in those with gastric corpus diseases. Among the 3805 serum samples matched with the H. pylori infection data, the serum gastrin-17 level was higher in the H. pylori-positive group than in the H. pylori-negative group. CONCLUSIONS: In people over 60 years of age, the serum gastrin-17 level tends to increase. In subjects with precancerous gastric lesions, it may increase significantly with the progression of gastric disease, and ultimately decrease in gastric cancer. Serum gastrin-17 is a good biomarker to differentiate benign from malignant gastric diseases. The site of the gastric lesions is an important factor affecting the serum gastrin-17 level, whereas H. pylori infection is usually associated with its increment.

[A molecular epidemiological study on the relationship between the polymorphism of GSTP1 and susceptibility to gastric cancer in northern Chinese].

We analyzed the distribution of glutathione-S-transferase P1 gene (GSTP1) polymorphism in a population from northern China and the relationship between the polymorphic BsmAI site in its exon 5 and gastric cancer susceptibility and evaluated the combined effect of GSTP1 polymorphism and H. pylori infection on gastric cancer. Blood samples were taken from 1,612 subjects in areas of high and low incidence of gastric cancer. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were performed to analyze the genotype of a GSTP1 polymorphism in exon 5 (Ile105Val). Serum levels of anti-H. pylori IgG were measured by enzymed-linked immunosorbent assay. We found that the GSTP1 Val variant allele frequency was 22%, which was significantly different from the Western people. There was a significant difference in GSTP1 allele gene distribution between the area of high incidence of gastric cancer(23%) and low incidence(20%). The frequency of GSTP1 Val/Val genotype was statistically higher in the gastric cancer group compared to the non-gastric cancer population. Analysis showed a statistically significant 1.587-fold increase in gastric cancer risk associated with the GSTP1 Val allele. Moreover, there was a statistically significant interaction (odds ratio, 17.571; 95% confidence interval, 6.207 - 49.742) between GSTP1 Val/Val genotype and positive H. pylori IgG status. Our results indicate that the distribution of GSTP1 polymorphism has geographic differences. Individuals with the GSTP1 Val allele gene show an increased risk for gastric cancer. Association of the GSTP1 (Val/Val) genotype with H. pylori IgG positive status could significantly increase gastric cancer risk.

[Effect of luteolin on COX-2 and mPGES-1 expression in LPS-induced RAW264.7 cells].

OBJECTIVE: To study the effect of luteolin on COX-2 and mPGES-1 expression in LPS-induced RAW264.7 cells. METHODS: After being pretreated with different concentrations of luteolin for 30 min, and then incubated with 1 microg/ml LPS for 12h, the effect of luteolin on the product of PGE2 in RAW264.7 cells was measured by enzyme immunoassay (EIA). The mRNA expression of COX-2 and mPGES-1 in RAW264.7 cells were analysed by RT-PCR. The COX-2 and mPGES-1 protein expression in RAW264. 7 cells were analysed by western blotting. RESULTS: Luteolin inhibited the LPS-induced PGE, synthesis in RAW264.7 cells. The mRNA and protein expression of COX-2 and mPGES-1 in RAW264.7 cells were also decreased by luteolin. CONCLUSION: Luteolin can inhibit significantly the expression of COX-2 and mPGES-1 in PGE2 synthetic pathway.

[Induction of apoptosis in human nasopharyngeal carcinoma cell line CNE-2Z by curcumin].

BACKGROUND & OBJECTIVE: Nasopharyngeal carcinoma (NPC) is a common malignant tumor in southern China. Radiotherapy is a major therapy for NPC in early phase,while proper chemotherapy is necessary for NPC in late phase. Some chemotherapeutic drugs have been used to treat tumors by inducing apoptosis of tumor cells. This study was to investigate effects of curcumin on proliferation,and apoptosis of NPC cell line CNE-2Z. METHODS: CNE-2Z cells were treated with different concentrations of curcumin, inhibition rates of cell proliferation,and IC(50) of curcumin were detected by MTT method. Cell apoptosis was analyzed by flow cytometry (FCM), agarose gel electrophoresis,and Hoechst33258-PI fluorescence staining. RESULTS: Proliferation of CNE-2Z cells was inhibited obviously by curcumin in dose- and time-dependent manners. IC(50) of curcumin to CNE-2Z cells at 24,48,and 72 h were (24.05+/-0.47),(19.20+/-0.17),and (7.35+/-0.50) micromol/L, respectively. After treated with 5,10,and 20 micromol/L of curcumin for 24 h, apoptotic rates of CNE-2Z cells were (4.9+/-3.2)%,(10.7+/-2.7)%, and (14.7+/-0.5)%, respectively. After treated with 10,20 micromol/L of curcumin for 24 h, morphologic changes, such as chromatin shrinkage,nuclear condensation,and chromatin fragmentation,were observed in CNE-2Z cells by fluorescent staining, a fragmented DNA ladder was detected by electrophoresis. CONCLUSION: Curcumin may induce apoptosis, and inhibit proliferation of CNE-2Z cells.