Activated sludge is a complicated microbial ecosystem that has diversity. Based on the diversity,microorganisms are selected by acclimation condition:the survival of the fittest,otherwise eliminate through selection or contest. Moreover,microorganisms in activated sludge acclimatize themselves to acclimation conditions. In the process the ecosystem distributes and adjusts microbial niche over again. The theory of "selection and acclimatization" may be used to explain the mechanism of activated sludge acclimation. So then,original sludge,wastewater quality and treating process are major influencing factors for activated sludge acclimation.

This study aims to analyse the value of CODEHOP RT-PCR in the detection of Flavivirus. According to the amino acid sequences of polyproteins of different flaviviruses published in GenBank, a pair of primers was designed using the CODEHOP method. One-step RT-PCR was used to detect Japanese encephalitis virus strain JEV1201, Dengue virus strain JKD001, and yellow fever virus vaccine YV6161. BLAST analysis and phylogenetic analysis were performed after the RT-PCR products of nucleocapsid genes were sequenced. The results showed that this method could amplify Flavivirus specifically, and the size and sequence of the target fragment accorded with the anticipated result. JEV1201 had the highest homology to Japanese encephalitis virus strain YL2009-4/YC2009-3, belonging to the branch of the phylogenetic tree of Japanese encephalitis virus strains. JKD001 had the highest homology to Dengue virus strain DENV-2/ID/1022DN/1975, belonging to the branch of the phylogenetic tree of Dengue virus strains. YV6161 had the highest homology to Yellow fever virus strain 17D, belonging to the branch of the phylogenetic tree of Yellow fever virus strains. In conclusion, the method of CODEHOP RT-PCR can be effectively used to detect, identify, and phylogenetically analyse Flavivirus.
DNA Primers ; genetics ; Flavivirus ; classification ; genetics ; isolation & purification ; Flavivirus Infections ; virology ; Humans ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Nonstructural Proteins ; genetics

OBJECTIVETo develop a recombinant vaccinia virus vaccine expressing HPV58 E7 and to determine its immuno-protective activity in mice bearing HPV58 E7+ tumor.

METHODSE7 DNA was amplified and cloned from a plasmid containing HPV58 E7 genome by PCR. To abolish its transforming activity, the nucleotides coding for amino acid residues at positions 24 and 92 were modified by site-directed mutagenesis so that cysteine was substituted by glycine. Balb/c 3T3 cells were transfected with mE7. The expression of E7 protein by the mE7-transfected Balb/c cells was confirmed by immunofluorescence staining. The transfected cells were observed in vitro for anchorage-independent growth and tumorigenesis in nude mice. Recombinant E7 vaccinia virus vaccine was constructed by homologous recombination of HPV58 E7 vaccinia expression plasmid and vaccinia virus (Tiantan stain). The immuno-protective activity of the vaccines was determined by tumor growth inhibition and cytotoxic T lymphocytes (CTL) induction in vaccine-immunized syngeneic mice.

RESULTSSubstitution of cysteine by glycine at both positions 24 and 92 of HPV58 E7 abolished its transforming activity. Growth of HPV E7+ tumor in mice immunized with the recombinant vaccinia virus expressing HPV58 E7 was inhibited, and the surviving time of the immunized mice was prolonged. CTL activity was induced as revealed by in vitro cytotoxicity assay using E7+ tumor cells as target cells.

CONCLUSIONSHPV58 E7, with its transforming potential abolished, may be used as vaccine for immunotherapy of patients with HPV 58 related cancers.

Animals ; Mice ; Mice, Inbred BALB C ; Mutagenesis, Site-Directed ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, Synthetic ; immunology ; Vaccinia virus ; genetics ; immunology

Objective To observe the Lactulose Oral Solution's influence in PPH postoperative complications and patients′ satisfactory rate for constipation and to find more intervention measures for reducing the complications. Methods A hundred and twenty cases of hemorrhoids with constipation were randomly divided into the two groups: the experimental group, the control group in 60 cases in each group. All patients’ postoperative bleeding, anal edema, stool impaction and satisfaction were observed and compared within 30 days. Results In the control group, the postoperative bleeding’ cases were of 23, anal edema’s were of 20 and stool impaction‘s were of 8. The incidence was 38.3%, 33.3% and 13.3% severally. The satisfactory patients were of 49 and the degree of satisfaction was of 81.7%. In the experimental group, the postoperative bleeding’ cases were of 12, anal edema’s were of 8 and stool impaction's was of 1. The incidence was 20%, 13.3% and 1.7% respectively. The cases who were satisfactory were of 58 and the degree of satisfaction was 96.7%. The postoperative bleeding , anal edema , stool impaction and satisfaction were different statistically in the two groups above (P < 0.05). Conclusion It can reduce PPH postoperative complications and increase patients′satisfaction by using Lactulose Oral Solution in patients with constipation and it will have a certain clinical value if these patients take this oral solution routinely.

Objective To observe the clinical effect of thrombus removal of external hemorrhoid combined with procedure for prolapse and hemorrhoids(PPH) in the treatment of circumferential mixed hemorrhoid with incarceration.Methods A total of 118 cases of circumferential mixed hemorrhoid with incarceration were divided into two groups:experimental group of 60 cases was treated by thrombus removal combined with PPH;control group of 58 cases was treated by Milligan-Morgan.We observed the differences of postoperative visual analogue scale (VAS)score,edema,bleeding,residual skin tag,wound healing time,anal stenosis,fecal incontinence,anorectal manometry and satisfaction in the two groups.Results There was significant difference between the experimental group and the control group in postoperative edema,bleeding and residual skin tag(χ2=6.63,4.19,6.64,P<0.05),but postoperative anal stenosis and fecal incontinence weren′t statistically significant different(χ2=0.38,1.11,P>0.05).Postoperative VAS score,wound healing time,satisfaction,anal resting and anal maximal squeeze pressure between the two groups were all statistically different(P<0.01).Conclusion The operation of thrombus removal of external hemorrhoid combined with PPH can effectively reduce the postoperative complications and promote recovery.

Objective To cluster analysis the population pharmacokinet-ic characteristics of nifedipine.Methods The data of mean pharmacoki-netic parameters and other factors may affecting the pharmacokinetics of drug were collected, normalized, and compared by multivariate cluster a-nalysis procedure using SAS software.Results The pharmacokinetics of Chinese Han population was similar to that of east Asia populations clas-sified as a subgroup, with AUC was (292.53 ±8.01) ng· mL-1 · h, tmax was (0.61 ±0.03) h, Cmax was (140.01 ±11.38) ng· mL-1 and t1/2 was (3.27 ±0.50) h, respectively.Conclusion The meta-analy-sis identified ethnic factor as a statistically significant factor of pharmaco-kinetics, subgroup of people located with Chinese Han population should reduce the dose of formulation.

OBJECTIVETo investigate the clinicopathological features, histogenesis and prognosis of mucinous tubular and spindle cell carcinoma (MTSCC).

METHODSFive MTSCCs were studied with histochemical, immunohistochemical staining, electron microscopy, and review of the related literatures.

RESULTSFour cases of MTSCC were females and one was male. Three patients presented with flank discomfort and two were incidentally found with health examination. In gross examination, the tumors were circumscribed. The cut surface was solid, gray-white, yellow or red. Necrosis was present in one case of high-grade MTSCC. Microscopically, low-grade MTSCC was mainly consisted of tubular, spindle cell and mucinous stroma with relatively bland morphology, and mitoses were rare. While in the high-grade area of one case, the cells were spindle or polymorphic with severe atypia and high mitotic activity, without mucinous stroma and tubular structure. Mucin was positive for Alcian blue. The neoplastic cells were positive for vimentin (5/5), CKpan (5/5), CK7 (5/5), CK19 (5/5), 34betaE12 (1/5), EMA (5/5), E-cadherin (3/5), CD10 (1/5), P504S (5/5), and CAM5.2 (5/5). The Ki-67 index was low (< or = 5%) in the low-grade component, while it was high (15%) in the high-grade component. Ultrastructural study showed short microvilli along glandular lumens. The nuclear membrane was focally invaginated. Four cases were followed up for 3 to 52 months, and recurrence and metastasis were not found.

CONCLUSIONSMTSCC occurs predominantly in females and it is a rare kidney neoplasm. Most of MTSCCs are low-grade and the prognosis is relatively good. However, the patients of high-grade MTSCC should be closely followed up.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; surgery ; Adult ; Carcinoma ; metabolism ; pathology ; surgery ; Carcinoma, Medullary ; pathology ; Carcinoma, Renal Cell ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Keratins ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Leiomyosarcoma ; metabolism ; pathology ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Nephrectomy ; Racemases and Epimerases ; metabolism ; Vimentin ; metabolism

Apoptosis ; Caspase 3 ; Caspases ; physiology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Isotretinoin ; pharmacology ; Melanoma ; pathology ; Receptors, Retinoic Acid ; physiology ; Retinoid X Receptors ; physiology ; Tretinoin ; pharmacology

Streptomyces hygroscopicus 17997 produces the antiviral and antitumor ansamycin antibiotic, geldanamycin. Studies on geldanamycin biosynthetic pathway will provide good tools for genetic manipulation of the antibiotic-producing strain to improve the productivity or to facilitate making novel geldanamycin analogs. The structural similarities between geldanamycin and ansamycins such as rifamycin or ansatrienin suggest that both geldanamycin and ansamycins has a closely related pathways of biosynthesis and that biosynthetic system for geldanamycin is similar to the one of type I polyketide synthase (PKS) enzyme system. To explore the possible PKS genes involved in geldanamycin biosynthesis, the degenerate primers were designed according to the conserved sequence of KS-AT region from erythromycin and oleandomycin type I PKS genes. Cosmids containing multiple PKS genes (pCGBK2,4,6,10,11,18) were obtained by hybridization with the PCR products, which were amplified from S. hygroscopisus 17997 genomic DNA. The designed primers above were used for PCR. Development of a Streptomyces temperate phage phiC31-derivative KC515( tsrR) transduction system was carried out for identification of cosmids containing the PKS gene related to biosynthesis of geldanamycin. Several factors, mainly the Ca2+ and Mg + concentrations in different culture media affecting the frequency of gene transfection, were optimized .Transfection efficiency could reach up to 10(3) /microg DNA on YMG medium supplemented with 10mmol/L MgSO4. Reversely, the transfection efficiency decreased when YMG medium was supplemented with 30mmol/L MgSO4. Gene transfection system based on the integration-defective phage KC515 had been established for S. hygroscopicus17997. Recombinant phages (ph111, 258, 287, 116, 105) were constructed by insertion of the homologous to PKS gene fragments into the KC515 phage vector. Gene disruption experiments were performed by transduction of recombinant phages into S. hygroscopicus 17997 genome, and disruption of geldanamycin production was observed as a result of homologous recombination between the cloned insert in recombinant phage and the S. hygroscopicus 17997 genome by integration. Thiostrepton resistant transductants were selected and integration event was analyzed by Southern hybridization. The fermentation broth extracts from five resistant transductants were analyzed by TLC and HPLC. The results showed that only G16 mutant failed to produce geldanamycin. This result showed that the integration of the insert DNA fragment in recombinant phage phl6 into the chromosome of S. hygroscopicus disrupted the expression of the geldanamycin biosynthetic genes. The original cosmid pCGBK10 containing this cloned insert was predicted to encode PKS genes in the geldanamycin biosynthesis. This study laid the foundation for cloning the PKS genes involved in geldanamycin biosynthetic gene cluster from S. hygroscopicus 17997.
Alcohol Oxidoreductases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Bacteriophages ; genetics ; Benzoquinones ; metabolism ; Blotting, Southern ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Genetic Vectors ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; genetics ; physiology ; Polymerase Chain Reaction ; Streptomyces ; genetics ; metabolism