Some important network addresses of microbiology teaching reso ur ces and two effective searching engines in Internet were provided. How to searc h, download and apply these resources to the teaching practice were discussed in this paper.

Objective To investigate the effects of gonadotropin releasing hormone agonist (GnRH- a) and antagonist (GnRH-ant) on cyclophosphamide (CTX)-induced ovarian damage in rats.Methods Seventy-two Sprague-Dawley female rats were divided randomly into six groups,which received normal saline (NS),CTX,GnRH-a+NS,GnRH-a+CTX,GnRH-ant+NS,and GnRH-ant+CTX respectively.Levels of serum follicle-stimulating hormone (FSH),luteinizing hormone (LH) and estradiol (E_2) were measured successively by the enzyme-linked immunosorbent assay method,and half of the rats were killed in the first week and between the fourth and the fifth week after stop of medication,respectively to compare the weight of the ovaries and the number of the primordial follicles and the growth follicles.Results (1) Throughout experiment,the serum levels of FSH,LH and E_2 of the control group fluctuated slightly,while those in the CTX group kept rising.During medication treatment,compared with the control group[(118?16) ?g/L, (350?35) ?g/L] and the CTX group[(113?15) ?g/L,(289?42) ?g/L],the concentrations of LH [(42 ?8)-(47?7) ?g/L,(31?5)-(36?7) ?g/L] and FSH [(124?45)-(136?32)?g/L,(178 ?54)-(198+27)?g/L] in the GnRH-a groups and the GnRH-ant groups were maintained at low levels significantly and the levels of LH in the GnRH-ant groups were significantly lower than that in the GnRH-a groups,but the levels of FSH in the GnRH-ant groups were significantly higher than that in the GnRH-a groups(P0.05),but the levels of FSH,LH and E_2 of the GnRH-ant+CTX group rose obviously and were similar to the levels of the CTX group,especially the FSH,and the levels of LH and FSH of the GnRH- ant + CTX group [(156?12) ?g/L,(520?44) ?g/L] and the CTX group [(178?18) ?g/L,(546?36) ?g/L] were significantly higher than that of the other four groups [(121?15)-(132?13) ?g/L,(335 ?35)-(359?26) ?g/L] at the 4~(th)-5~(th) week after stop of treatment(P0.05),but the number of all kinds of follicles declined significantly in the GnRH-ant+CTX group[(195?15),(36?12)] and the CTX group [(212?11),(36?9)] compared to the other four groups[(302?15)-(690?43),(44?12)-(58?11),P

Objective To study the genetic variations between measles vaccinc strain S191 and strains that circulated in Zhejiang province causing the epidemics during 1999 to 20 1 1.Methods Complete sequence of the nine Zhejiang measles strains were amplified by RT-PCR assay.Products were sequenced and the obtained sequences were aligned and analyzed with vaccine strains S191 and the major epidemic strains isolated in foreign countries.Results The homology of amino acid among the nine Zhcjiang strains were 98.77％-99.89％.The strains were not affected by positive selection and the variations on each gene were still in random drift.Compared to vaccine strain S191,there were 135 to 159 amino acid changes in Zhejiang measles virus,in which 113 points were common variable positions,resulting in mutations on five glycosylation sites.At the nucleotide level,the biggest differences between the Zhejiang strains and the vaccine strain S191 were found on N gcne,with the average divergent ratio as 5.5％,while the biggest one was P protein,in the amino acid level,with the average mutation rate as 7.7％.In addition,with the complete genome sequences,the genetic distance between Zhcjiang epidemic strains and vaccine strains was greater than the distances between epidenic strains of genotype D4,B3 and vaccine strains (t=-9.76,P＜0.05；t=-12.39,P＜0.05).Conclusion There were significant differcnccs found in the each of the genes between Zhejiang epidemic strains and the vaccine strain S191.The differences between the current vaccine strains and H genotypc epidemic strains were much larger than the differences between the vaccine and the forcign epidemic strains (genotype D4,B3).Therefore,wc should pay close attcntion to this trend,and to develop candidates for the dcvclopnent of vaccines,as early as possible.

Objective To study the evolutionary characteristics and rules of two lineages on influenza B virus.Methods A total of 126 HA1 sequences of strains isolated during 1940 to 2012were downloaded from the GenBank.Time of the most recent common ancestor (TMRCA) and divergence of the two lineages were calculated based on the data from phylogenetic analysis of HA1gene,using Bayesian Markov Chain Monte Carlo (Bayesian-MCMC) and molecular clock method.Results The average amino acid variant ratios were ranged from 5.4％ to 10.2％ within the strains of influenza B virus isolated during 1978 to 2010.Compared with the Victoria-like strains,all Yamagatalike strains showed an amino acid deletion at 163th site,while some of them showing a deletion at position 166.HA1 gene of influenza B virus seemed not have been affected by positive selection except a few sites.The evolutionary average rate on HA1 gene was 2.138 × 10-3 substitutions/site/year (95％HPD:1.833 × 10-3-2.437 × 10-3 substitutions/site/year).The estimated dates for TMRCA of the two lineages of influenza B virus could be dated back to 1971 (95％ HPD:1969-1972),while the divergence times of the two lineages were 1973 (95％ HPD:1971-1974) and 1977 (95％ HPD:1975-1978) respectively.Conclusion Significant differences were found on HA1 gene between earlier and recent identified strains of Victoria and Yamagata lineage.Differences between the two lineages increased and showing the potential of dividing themselves into different subtypes in the future.More attention should be paid to these trends and the related epidemiological significance.

Objective To analyze the genetic characteristics of the complete sequence of coxsackievirus A24 variant(CA24v) isolated from acute hemorrhagic conjunctivitis (AHC) outbreaks in Zhejiang province during 2002 to 2010.Methods Complete sequences of CA24v epidemic strains isolated in different years were amplified under the RT-PCR assay,while the sequences of whole genome,VP1,and 3C region of Zhejiang strains were compared with epidemic strains isolated in other areas of China and abroad.Results The whole genome of Zhejiang CA24v strains isolated in 2002 and 2010 was 7456-7458 bp in length,encoding a polyglutamine protein which containing 2214 amino acid residues.There was a insertion with T on site 97 and 119 within 5' non-coding region between epidemic strain Zhejiang/08/10 and strains isolated in 2002.The rates of amino acid homology among Zhejiang/08/10 and other strains isolated since 2002 were between 94.7％ and 100.0％.Compared with the representative strains circulated within the recent 60 years,the largest average amino acid variations had been occurred on region 2A and 3A,with the ratios as 8.4％ and 7.3％ respectively.The smallest variation happened in region 3D,with the ratio only as 1.9％.The rates of stable amino acid variation on the whole genome between strains isolated since 1987 and 2002 were 38 and 20.P-distance within groups appeared that region 3C was more stable than VP1 of strains isolated in 2002-2010,and the 3D of early strain Jamaica/10628/87 might have had a nature of recombination but not observed on those epidemic strains in recent years.Conclusion Within the evolution of CA24v strains,the time course was more significant than the geographical differences.There had been sporadic epidemics of AHC caused by CA24v in Zhejiang province since 2002.

OBJECTIVETo investigate the nephrotoxocity of Longdan Xiegan Tang in rats, and to test its safety.

METHODSprague-Dauley rats were given Longdan Xiegan Tang 4.5 mL x (100 mg)(-1) Bid for thirty days, and the control group was given NS. MTP, BUN, Cr, MDA, MTP/Ucr and SOD were measured on 1st, 2nd, 3rd, 4th week. The kidney tissues were viewed with light microscopy and electron microscope.

RESULTMTP and MTP/Ucr were obviously higher than controls ( P < 0.01), and the other index had no difference (P > 0.05). No remarkable structural change could been seen with light microscopy, but with electron microscope we could find that the basal membranes were thickened and some of foot process were infused.

CONCLUSIONLongdan Xiegan Tang will result in injury of kidney function.

Animals ; Basement Membrane ; ultrastructure ; Blood Urea Nitrogen ; Creatinine ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Gentiana ; chemistry ; Kidney ; ultrastructure ; Kidney Cortex ; metabolism ; Magnoliopsida ; chemistry ; Male ; Malondialdehyde ; metabolism ; Plants, Medicinal ; chemistry ; Proteinuria ; chemically induced ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Two distinct routes (classical mevalonate pathway and a novel mevalonate-independent pathway) are utilized by plants for the biosynthesis of isopentenyl diphosphate, the universal precursor of isoprenoids (Fig. 1). Present researches indicated that taxol was synthesized mainly via non-mevalonate pathway, but not genetic evidence was showed. The second step in non-mevalonate pathway involves an intramolecular rearrangement and subsequent reduction of deoxyxylulose phosphate to yield 2-C-methyl-D-erythritol-4-phosphate, and 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) with responsibility for this reaction was considered as a key enzyme. As a tool for the isolation of genes in terpenoid biosynthesis in plants, total RNA was prepared from Taxus chinensis suspension cells, a cell type highly specialized for diterpene (taxol). A reverse transcription-PCR strategy based on the design of degenerated oligonucleotides was developed for isolating the gene encoding a gymnosperm homolog of this enzyme from Taxus chinensis. Through sequence analysis by Blast P online, the resulting cDNA showed highly homologous to 1-deoxy-D-xylulose 5-phosphate reductoisomerases, with 95% identification compared with Arabidopsis thaliana (Q9XFS9), 94% with Mentha x piperita (Q9XESO), 80% with Synechococcus elongatus (Q8DK30), 78% with Synechocystis sp. PCC 6803 (Q55663) and Nostoc sp. PCC 7120 (Q8YP49), and 73% with Synechococcus leopoliensis (Q9RKT1). Deduced amino acid sequences were also analyzed by PROSITE, ClustalX (1.81) and Phylio (3.6 alpha), and data present evidence for the existence of this deoxyxyluose phosphate reductoisomerase in Taxus chinensis. This is the first report of the dxr gene cloned from gymnosperm.
Aldose-Ketose Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; Mevalonic Acid ; metabolism ; Multienzyme Complexes ; genetics ; Oxidoreductases ; genetics ; Phylogeny ; RNA ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Taxus ; genetics

OBJECTIVETo investigate the effect of huanglian jiedu tang on Alzheimer' s disease, and its influence on cytokines.

METHODAD rats models were made by A beta injection, and then water-maze tests were made to investigate the study and memory ability. TNF-alpha, INF-gamma and IL-2 were examined by ELISA. After treatment with huanglian jiedu tang, the rats were evaluated again.

RESULTAfter A beta injection, the study and memory ability of the rats was decreased. By the treatment with huanglian jiedu tang, it was improved (P < 0.01); TNF-alpha, INF-gamma and IL-2 levels of AD rats were higher than those in control group (P < 0.01); after treatment the levels came down (P < 0.01).

CONCLUSIONHuanglian jiedu tang can improve the study and memory ability of AD rats by changing their immune state.

Alzheimer Disease ; blood ; psychology ; Animals ; Coptis ; chemistry ; Cytokines ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gardenia ; chemistry ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Male ; Memory ; drug effects ; Phellodendron ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Scutellaria ; chemistry ; Tumor Necrosis Factor-alpha ; metabolism

Objective: Autoimmune polyendocrine syndrome type Ⅰ(APS-Ⅰ) is caused by mutations in the autoimmune regulator gene (AIRE) gene. In this study, phenotype and AIRE gene analysis were performed in two patients with APS-Ⅰ. Methods: Peripheral blood samples were collected from two patients with APS-Ⅰ and their families. All exons of the AIRE gene and adjacent exon-intron sequences were amplified by PCR and subsequently sequenced. The silico analysis was performed to predict the possible impact of the mutations on the function of the AIRE protein. At the same time, 100 healthy controls were selected to confirm the mutation. Results: Case 1 was a 31-year-old female who exhibited chronic mucocutaneous candidiasis, hypoparathyroidism, Addison′s disease, Hashimoto′s thyroiditis, and premature ovarian failure. A homozygous c. 483_484insC mutation in exon 4 of AIRE gene was identified in this patient. Her parents, siblings and son were heterozygous for this mutation, which is consistent with the autosomal recessive inheritance pattern. Case 2 was a 34-year-old male who had mucocutaneous candidiasis, Addison′s disease, primary hypoparathyroidism, and Hashimoto′s thyroiditis. A compound heterozygous AIRE mutation (c.179A>G/C.463+ 2T>C) were identified in this patient. His father was heterozygous for c. 179A>G mutation, and his mother was heterozygous for C. 463+ 2T>C, which is consistent with autosomal recessive inheritance mode. The c. 483_484insC and c. 463+ 2T>C have been reported to be pathogenic. The c. 179A>G mutation was predicted pathogenic by SIFT and PolyPhen2 software, which was not detected in 100 healthy controls. It has not been reported in the HGDM database and is a novel mutation. Conclusion We identified a novel AIRE gene mutation (c.179A>G), which contributed to further understanding of the pathogenesis of APS-Ⅰ. The clinical variation and rarity of APS-Ⅰ makes the syndrome hard to recognize. Early recognition of symptoms and screening for AIRE mutation in patients with APS-Ⅰ has important clinical implications for the diagnosis and treatment.

The objective of study was to investigate the in vitro suppressive effect of angelica polysaccharide (APS) on human cytomegalovirus-induced apoptosis via direct infection in CHRF-288-11 cells. HCMV AD169 directly infected CHRF-288-11 were cultured in vitro, APS in different doses were added on day 3 after the infection of virus. Cells of every group were collected at different time points. HCMV DNA of cells were detected by using polymerase chain reaction and the apoptotic cells were examined by using Hoechst staining, MTT assay, DNA fragmentation assay and flow cytometry. The results showed that the APS to some extent inhibited the apoptosis of CHRF cells infected by HCMV in vitro in a dose-dependent manner. The expression of HCMV IEA in CHRF-288-11 cells was found by PCR amplification. Morphology observation, flow cytometry assay and DNA fragmentation assay revealed the existence of apoptosis. With the dose decrease of APS added to the infected CHRF cells, the percentage of apoptotic cells increased. It is concluded that the HCMV AD169 can infect CHRF cells directly in vitro and decrease cell viability. HCMV AD169 infection increases the apoptosis of CHRF cells in time-dependent manner. When APS was added to the CHRF cells infected by HCMV AD169 in vitro, the viability of CHRF cells increase, which indicated that APS to some extent protects the CHRF cells infected by HCMV. APS suppresses the cytomegalovirus-induced apoptosis in CHRF cells directly infected in vitro in dose-dependent manner.
Angelica ; chemistry ; Apoptosis ; drug effects ; Cells, Cultured ; Cytomegalovirus ; Humans ; Megakaryocytes ; cytology ; drug effects ; virology ; Polysaccharides ; pharmacology