141例肺炎休克逆转分析

肺炎休克为常见的感染中毒性休克,据国内报告占同期中毒性休克病例的首位。休克时由于血液动力学的变化,临床又表现为两种不同类型,即所谓“冷休克”型及“热休克”型。前者由于微循环痉挛收缩,血管内径缩小,容积降低,静脉回流量减少,阻力增加,心脏负荷加重,心输出量降低。临床表现为心跳加快,皮肤苍白,四肢冷湿,脉压缩小,眼底动脉痉挛;而后者则因微血管扩张,短 表1

急性白血病患者中医证型与hTERT mRNA表达相关性的研究

目的:探讨急性白血病患者hTERT mRNA表达的中医本质。方法:采用半定量RT-PCR法检测32例不同证型急性白血病患者外周血单个核细胞hTERT mRNA表达的异同。结果:急性白血病患者外周血hTERT mRNA表达明显高于正常人(P<0.01);急性白血病患者hTERT mRNA表达量由低到高依次为,瘀血痰结型<气阴两虚型<毒热炽盛型。其中毒热炽盛型和瘀血痰结型对比差别无统计学意义(P<0.05)。结论:hTERT mRNA的高表达可能反映了白血病毒热炽盛的本质。

Cloning,characterization,and expression analysis of the DEAD-box family genes,Fc-vasa and Fc-PL10a,in Chinese shrimp (Fenneropenaeus chinensis)

RNA helicases of the DEAD-box and related families are involved in various cellular processes including DNA replication, DNA repair, and RNA processing. However, the function of DEAD-box proteins in aquaculture species is poorly understood at molecular level. We obtained the full-length cDNA sequences of two genes encoding helicase-related proteins, Fc-vasa and Fc-PL10a, from the testes of Chinese shrimp, Fenneropenaeus chinensis. The two predicted amino acid sequences contain all the conserved motifs characterized by the DEAD-box family and several RGG repeats in the N-terminal regions. Homology and phylogenetic analyses indicate that they belong to the vasa and PLIO subfamilies. The three-dimensional structures of the two proteins were predicted with a homology modeling approach. Both core proteins consist of two tandem RecA-like domains similar to those of the DEAD-box RNA helicase. Using reverse transcription-polymerase chain reaction （RT-PCR） and real-time PCR we found that Fc-vasa was expressed specifically in the adult gonads. Transcription decreased in the ovary but increased in the testis during gonadal development. Fc-PL10a expression was widely distributed in the tissues we examined. Using in situ hybridization, we demonstrated that the Fc-vasa transcript is localized to the cytoplasm of the spermatogonia and oocytes. Thus, our results suggest that Fc-wasa plays an important role in germ-line development, and has utility as a germ cell lineage marker which will help to generate new insight into the origin and differentiation of germ cells as well as the regulation of reproduction in F. chinensis.

Cloning and Sequence Analysis of Genome from the Inner Mongolia Strain of the Endogenous Betaretroviruses (enJSRV)

In order to amplify the complete genome of enJSRV from the strain of Inner Mongolia (enJSRV-NM), we used enJSRV-specific and JSRV-specific DNA probes in dot blot hybridization. Seven pairs of primers were designed based on Genbank sequences. Seven fragments were obtained by PCR and were cloned into the PMD19-T vectors. The recombinant plasmids were sequenced and analyzed. The results showed that the genome was 7 942 bp in length and contained four overlapping open reading frames corresponding to the gag, pro, pol and env genes as well as an additional open reading frame (orf-x) that overlaps the 3' end of the pol gene. The nucleotide acid sequences of the enJSRV-NM loci were compared with the sequences of South Africa enJS56A1 strain (Accession No. AF153615) and USA JSRV21 strain (Accession No. AF105220). The nucleotide acid identities were 99.2% and 92.3% respectively. Two zinc fingers were found in the NC region in the predicted amino acid sequence. However, the YXXM motif, which is a reliable molecular marker for the infectious exogenous virus, was not found in the TM region. It was found that the enJSRV-NM region was 90%-98% identical at the amino acid level to its exogenous infectious counterparts in most of the retroviral genome. This is the first nucleotide sequence of enJSRV reported in P.R China. The resource work has provided a wide range of information useful not only for expression genomics and annotation of genomic DNA sequence, but also for further research on the clinical diagnosis of OPA.

Expression and Immunological Analysis of Capsid Protein Precursor of Swine Vesicular Disease Virus HK/70

VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore, anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection.

Indinavir Resistance Evolution in One Human Immunodeficiency Virus Type 1 Infected Patient Revealed by Single-Genome Amplification

Human Immunodeficiency Virus Type 1 exists in vivo as quasispecies, and one of the genome's characteristics is its diversity. During the antiretroviral therapy, drug resistance is the main obstacle to effective viral prevention. Understanding the molecular evolution process is fundamental to analyze the mechanism of drug resistance and develop a strategy to minimize resistance. Objective: The molecular evolution of drug resistance of one patient who had received reverse transcriptase inhibitors for a long time and had treatment which replaced Nevirapine with Indinavir was analyzed, with the aim of observing the drug resistance evolution pathway. Methods: The patient, XLF, was followed-up for six successive times. The viral populations were amplified and sequenced by single-genome amplification. All the sequences were submitted to the Stanford HIV Drug Resistance Database for the analysis of genotypic drug resistance. Results: 149 entire protease and 171 entire reverse transcriptase sequences were obtained from these samples, and all sequences were identified as subtype B. Before the patient received Indinavir, the viral population only had some polymorphisms in the protease sequences. After the patient began Indinavir treatment, the variants carrying polymorphisms declined while variants carrying the secondary mutation G73S gained the advantage. As therapy was prolonged, G73S was combined with M46I/L90M to form a resistance pattern M46I/G73S/L90M, which then became the dominant population. 97.9% of variants had the M46I/G73S/L90M pattern at XLF6. During the emergence of protease inhibitors resistance, reverse transcriptase inhibitors resistance maintained high levels. Conclusion: Indinavirresistance evolution was observed by single-genome amplification. During the course of changing the regimen to incorporate Indinavir, the G73S mutation occurred and was combined with M46I/L90M.

15-PGDH is reduced and induces apoptosis and cell cycle arrest in gastric carcinoma

AIM: To investigate the expression of 15-hydroxyprostaglandin dehydrogenase (15-PGDH) in human gastric cancer and it's mechanism in apoptosis and cell cycle arrest. METHODS: Expression of 15-PGDH mRNA and protein was examined by immunohistochemistry, immunocytochemistry, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting in tissue from human gastric cancer, gastric precancerous state (gastric polyps and atrophic gastritis), normal stomach, and gastric cancer cell lines. The relationship between gastric cancer, gastric precancerous state and 15-PGDH expression was determined. The association between expression of 15-PGDH and various clinicopathological parameters in gastric cancer was evaluated. Human gastric cancer cell line SGC-7901 was transfected with 15-PGDH expression plasmids. The effect of 15-PGDH on the cell cycle was examined by flow cytometry. The effect of 15-PGDH on apoptosis was examined by transmission electron microscopy, flow cytometry and transferasemediated nick end labeling (TUNEL) assay. Expression of cell cycle (p21, p27, p16 and p53) and apoptosis (Survivin, BCL-2, BCL-XL, BAK and BAX) genes was analyzed by RT-PCR. RESULTS: Expression of 15-PGDH mRNA and protein in human gastric cancer tissues was significantly lower than in normal gastric tissues (P < 0.01). Expression in human gastric cancer cell lines MKN-28 and MKN-45 was reduced, and absent in SGC-7901 cells (P < 0.05). Reduction of 15-PGDH expression was also found in precancerous tissues, such as gastric polyps and atrophic gastritis (P < 0.01). There was a significant difference in expression of 15-PGDH among various gastric cancer pathological types (P < 0.05), with or without distant metastasis (P < 0.05) and different TNM stage (P < 0.01). Flow cytometry demonstrated a significant increase in apoptotic cells in SGC-7901 cells transfected with pcDNA3/15-PGDH plasmid for 24 and 48 h (P < 0.01), and an increased fraction of sub-G1 phase after transfection (P < 0.05). TUNEL assay showed an increased Apoptotic Index in cells overexpressing 15-PGDH (P < 0.01). After transfection, expression of proapoptotic genes, such as BAK (P < 0.05), BAX and p53 (P < 0.01), was increased. Expression of antiapoptotic genes was decreased, such as Survivin, BCL-2 and BCL-XL (P < 0.01). Expression of cyclin-dependent kinase inhibitors p21 and p16 (P < 0.01) was significantly upregulated in cells overexpressing 15-PGDH. CONCLUSION: Reduction of 15-PGDH is associated with carcinogenesis and development of gastric carcinoma. 15-PGDH induces apoptosis and cell cycle arrest in SGC-7901 cells.

SEDIMENTARY-STRUCTURAL EVOLUTION OF A STRIKE-SLIP COAL BASIN IN CENTRAL YUNNAN,CHINA

The Kebao coal basin is located at the middle segment of the Xiaojiang fault zone in the central part of Yunnan Province. The coal-bearing strata of Tertiary age are made up of alluvial fan-lacustrine-swamp genetic sequences. Sinistral strike-slip motion of the N-S trending fault zone has created a locally transtensional or transpres sional tectonic environment, giving rise to the relative movement of fault blocks in the basement, and the development of a strike-slip coal basin at the surface. Based on de tailed sedimentary-structural analyses, the paper proposes geodynamic models for the coal basin formation and its deformation.

Construction and Genetic Analysis of Murine Hepatitis Virus Strain A59 Nsp16 Temperature Sensitive Mutant and the Revertant Virus

Coronaviruses (CoVs) are generally associated with respiratory and enteric infections and have long been recognized as important pathogens of livestock and companion animals. Mouse hepatitis virus (MHV) is a widely studied model system for Coronavirus replication and pathogenesis. In this study,we created a MHV-A59 temperature sensitive (ts) mutant Wu"-ts18(cd) using the recombinant vaccinia reverse genetics system. Virus replication assay in 17C1-1 cells showed the plaque phenotype and replication characterization of constructed Wu"-ts18(cd) were indistinguishable from the reported ts mutant Wu"-ts18. Then we cultured the ts mutant Wu"-ts18(cd) at non-permissive temperature 39.5°C,which "forced" the ts recombinant virus to use second-site mutation to revert from a ts to a non-ts phenotype. Sequence analysis showed most of the revertants had the same single amino acid mutation at Nsp16 position 43. The single amino acid mutation at Nsp16 position 76 or position 130 could also revert the ts mutant Wu"-ts18 (cd) to non-ts phenotype,an additional independent mutation in Nsp13 position 115 played an important role on plaque size. The results provided us with genetic information on the functional determinants of Nsp16. This allowed us to build up a more reasonable model of CoVs replication-transcription complex.

Na^+/K^+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress

Na^＋/K^＋-ATPases are membrane-associated enzymes responsible for the active transport of Na^＋ and K^＋ ions across cell membranes, generating chemical and electrical gradients. These enzymes＇ α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na^＋/K^＋-ATPase α-subunit eDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction （RT-PCR） and rapid amplification of eDNA end methods. Analysis of the nucleotide sequence revealed that the eDNA had a full-length of 3 833 base pairs （bp）, with an open reading frame of 3 120 bp, 5＇ untranslated region （UTR） of 317 bp, and 3＇ UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na^＋/K^＋-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of arnino acid sequences showed that the P. tritubereulatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit＇s transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab＇s Na^＋/K^＋-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.

XAF1 is frequently methylated in human esophageal cancer

AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction （MSP） in four esophageal cancer cell lines （KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines）, nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine （5-aza-dc）, a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS： MSP results were as follows： loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation （com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells）; all nine cases of normal esophageal mucosa were unmethylated; and 54/72 （75.00%） samples from patients with esophageal can- cer were methylated, and 25/72 （34.70%） matched adjacent tissues were methylated （75.00% vs 34,70%, z2 = 23.5840, P = 0.000）. mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue （Z2= 9.143, P = 0.002）. XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation （Fisher＇s exact test, P = 0.004）. Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION： XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.

解构分析与管理

解构分析,作为后现代认识论的代表,原本是一种文学评论方法,看上去好像与管理与组织无关。不过,这样的看法也许是错误的。通过促进这些颠覆意味文章的理解,通过对于文字之内二元对立的认知,通过对因果关系成因的逆转分析,解构分析为管理工作中的某些虚夸(rhetoric)提供了有效的批评工具。同时,解构分析本身并不在为自己创造它本意要否定的种种永久的等级层次,解构是个连续的过程,而文字的内容是不确定的,这就是解构分析所要阐述的本意。

Analyses of thermodynamic performance for the endoreversible Otto cycle with the concepts of entropy generation and entransy

In this paper, the endoreversible Otto cycle is analyzed with the entropy generation minimization and the entransy theory. The output power and the heat-work conversion efficiency are taken as the optimization objectives, and the relationships of the output power, the heat-work conversion efficiency, the entropy generation rate, the entropy generation numbers, the entransy loss rate, the entransy loss coefficient, the entransy dissipation rate and the entransy variation rate associated with work are discussed. The applicability of the entropy generation minimization and the entransy theory to the analyses is also analyzed. It is found that smaller entropy generation rate does not always lead to larger output power, while smaller entropy generation numbers do not always lead to larger heat-work conversion efficiency, either. In our calculations, both larger entransy loss rate and larger entransy variation rate associated with work correspond to larger output power, while larger entransy loss coefficient results in larger heat-work conversion efficiency. It is also found that the concept of entransy dissipation is not always suitable for the analyses because it was developed for heat transfer.

Artemisinin ameliorated proteinuria in rats with adriamycin-induced nephropathy through regulating nephrin and podocin expressions

OBJECTIVE:To investigate the effects of artemisinin against proteinuria and glomerular filtration barrier damage in rats with adriamycin-induced nephropathy,and the potential mechanism underpinned the action.METHODS:Forty adriamycin rats were randomly divided into two groups with the ratio of 1︰3;the small-number group served as control group(n=10),and the rats in the large-number group were treated with adriamycin to induce nephropathy;then they were further randomly assigned into 3subgroups:benazepril group(n=10),artemisiningroup(n=10),and adriamycin group(n=10).The benazepril group and artemisinin group were treated with benazepril suspl(5.0 mg/kg daily)and artemisinin suspl(150 mg/kg daily)respectively after being modeled;those in the control group and adriamycin group were intragastrically administered an equivalent volume of distilled water every day.The treatment after model establishment lasted for a total of 4 weeks.The 24 h uric protein,blood biochemicals,renal pathological changes,renal ultrastrutural changes,Nephrin and Podocin proteins and gene expressions were measured by Coomassie brilliant blue assay,completely automatic biochemical analyzer,light microscope,electron microscopy,Western blot and reverse transcription polymerase chain reaction,respectively.RESULTS:The rats in adriamycin group showed a significant increase in 24 h uric protein excretion,serum total cholesterol(TC),triglyceride(TG),blood urea nitrogen(BUN),serum creatinine(Scr)and decrease in albumin(Alb)(P<0.05 or P<0.01).Compared with adriamycin group,artemisinin could reduce uric protein excretion,decrease the serum TC,TG elevation,increase the serum Alb level,up-regulate the expressions of Nephrin and Podocin(P<0.05 or P<0.01),but no statistical significance effects on the levels of BUN,Scr in artemisinin group(P>0.05).The renal pathological and ultrastrutural observation indicate that artemisinin could attenuate the severity of foot process effacement and fusion in the nephropathic rats.CONCLUSION:Artemisinin might have an effect on the nephropathy in rats caused by adriamycin,which may be at least partly correlated with attenu-ation of the severity of foot process effacement and fusion,up-regulation of the expressions of Nephrin and Podocin in the glomeruli in the rats.

Effect of quercetin on secretion and gene expression of leptin in breast cancer

OBJECTIVE: To investigate the possible inhibitory action of pure quercetin on secretion and gene expression of leptin in the T47 D breast cancer cell line.METHODS: In this experimental study, T47 D cells were cultured in monolayers in RPMI 1640. IC50 was determined by MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay after 24 h treatment at different concentrations of quercetin.The levels of leptin gene expression were measured by reverse-transcription real-time polymerase chain reaction(PCR). Secreted leptin was measured in the supernatant of cells by an enzyme-linked immuno sorbent assay.RESULTS: Analysis of MTT assay data showed that quercetin has a cytotoxic effect on T47 D breast cancer cells with 40 μM IC50 after 24 h exposure. Dataanalysis of real-time PCR showed that with increases in quercetin concentration, a decreasing trend was seen in m RNA levels of leptin of treated cells compared with the control cells(P < 0.05). Also,measurement of secreted leptin in the culture media showed a similar decreasing trend in the amount of leptin protein in the treated cells compared with the control cells(P < 0.05).CONCLUSION: Quercetin significantly inhibits the growth of T47 D cells through inhibition of leptin secretion and gene expression in T47 D breast cancer cells. Therefore, it might be an alternative approach to breast cancer therapy through leptin targeting.