Fomentation engineering is one of the modern biotechnologies. It has been intensively studied and widely applied in edible fungi. Based on the review of research history of liquid fermentation for edible fungi, the research status of liquid fermentation about edible fungi were summarized, and its application prospects on edible fungi production of our country were described in this paper.

Constructing eukaryotic expressing vector of pMT/Bip/V5-His A-HIV gp120 and transfecting S2 cells to establish stable gp120-expressing S2 cell line. The gp120 gene of HIV CRF07-BC epidemic in China was amplified by polymerase chain reaction from the recombinant vector PRSV-gp120 and confirmed by DNA sequence analysis. The DNA fragment of HIV gp120 was digested with the restriction endonucleases NcoI and XhoI, then inserted into eukaryotic expressing vector pMT/BiP/V5-His A. A selection vector containing the streptomyces griseochromogenes bsd gene conferring blasticidin resistance was cotransfected into drosophila S2 cells by Cellfectin II reagent. The stable cell line was established following repeated screening by blasticidin. HIV gp120 was purified by a Ni-NTA affinity column followed by molecular sieve chromatography. Recombinant protein was characterized by SDS-PAGE, Western blot and ELISA. The eukaryotic expressing vector pMT/BiP/V5-His A-HIV gp120 was constructed, stable expressing cell line was established, and protein was expressed and purified successfully. This result will contribute to functional study of gp120 and vaccine design against AIDS.
Animals ; Cell Line ; Drosophila ; genetics ; metabolism ; virology ; Gene Expression ; HIV ; genetics ; metabolism ; HIV Envelope Protein gp120 ; genetics ; isolation & purification ; metabolism ; HIV Infections ; virology ; Humans ; Recombinant Proteins ; genetics ; isolation & purification ; metabolism

In the present study, the modulatory effect of AMPA receptors on gamma-aminobutyric acid (GABA) transporter current was investigated on enzymatically isolated horizontal cells of carp retina. The GABA transporter current elicited by 1 mmol/L GABA was decreased immediately after pre-application of AMPA (30 mumol/L or 3 mmol/L) for 50 s. Application of 10 mmol/L BAPTA in intracellular solution inhibited the suppression effect of AMPA on GABA transporter current. The suppression effect induced by co-application of 3 mmol/L AMPA and 3 mmol/L NMDA was similar to that of 3 mmol/L AMPA or 3 mmol/L NMDA alone. These results suggest that the activation of AMPA receptors inhibits GABA transporter-mediated current by affecting intracellular Ca(2+) processes in the retinal horizontal cells, which is identical with the modulatory effect of NMDA receptors on GABA transporters.
Animals ; Carps ; Egtazic Acid ; analogs & derivatives ; pharmacology ; GABA Plasma Membrane Transport Proteins ; metabolism ; Receptors, Ionotropic Glutamate ; metabolism ; Retinal Horizontal Cells ; metabolism ; gamma-Aminobutyric Acid ; pharmacology

Calcium is one of the most versatile intracellular second messengers, which plays crucial roles in many intracellular signaling pathways. Researches on intracellular calcium distribution, regulation and function are important for our understanding of cellular physiology. In this mini-review, the regulation of intracellular calcium signal in retinal horizontal cells and the relevant physiological functions were introduced based on the experiments carried out in our laboratory. Intracellular calcium dynamics following the activation of AMPA and NMDA receptors were introduced based on our experiments performed on carp retinal horizontal cells using calcium imaging technique and computational methods. An initial peak response was observed in both cases, which indicated an active participation of intracellular calcium store during the calcium dynamics initiated by AMPA/NMDA receptor activation. Intracellular recording experiments indicated that calcium signaling was crucial for the gradual enhancement of the retinal horizontal cell's responsiveness in exposure to repetitive red flashes. Possible roles of intracellular calcium signaling in the regulation of GABA transporter activity were also introduced based on our whole-cell recording experiments performed on isolated carp retinal horizontal cells.
Animals ; Calcium ; metabolism ; Calcium Signaling ; Carps ; Cells, Cultured ; Patch-Clamp Techniques ; Receptors, AMPA ; metabolism ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Retinal Horizontal Cells ; physiology

Diacylglycerol, DAG, because of its multifunctional and nutritional properties, attracted considerable attention recently. Enzymatic synthesis of diacylglycerols from linoleic acid was investigated in a solvent-free reaction in a continuously operated fixed bed reactors containing Lipozyme RM IM. By appropriate manipulation of the fluid-residence time, the relative proportions of the various acylglycerols in the effluent stream can be controlled. In addition, the presence of excess glycerol is effective for the removal of water produced during the esterification reactions. Under the conditions of molar ratio of linoleic acid to glycerol of 0.5, the immoblized enzyme maintained high stability and allowed the reaction to continue for 10 days without significant deterioration in enzyme activity. It was determined that the conversion of fatty acid, content of 1,3-DAG and volume efficiency of reactor reached optima under the conditions: a packaged-bed reactor(with a ratio of packed length to inner diameter of 7.8), reacting temperature at 65 degrees C, molar ratio of linoleic acid to glycerol of 0.5, and feeding flow rate of 1.2 mL/min.
Catalysis ; Diglycerides ; chemical synthesis ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; Lipase ; chemistry

OBJECTIVE: To explore the clinical features of acute urine retention (AUR) ocurring in patients with benign prostate hyperplasia (BPH). METHODS: Clinical data from 548 patients diagnosed with BPH were retrospectively studied, and the clinical parameters of these patients with or without AUR were analyzed by statistical methods. RESULTS: Development of AUR was found in 164 patients (29.9%). Patients' age, IPSS, maximum flow rate (Q(max)), residual urine volume, prostate volume, transition zone volume, prostate-specific antigen (PSA) density, total PSA (tPSA) and free PSA (fPSA) including the ratio of free to total PSA(f/tPSA) were significantly different in the 2 groups (P<0.05), while there were no significant differences in the disease duration and transition zone index between the 2 groups (P>0.05). Multivariate logistic regression analysis showed that IPSS score, residual urine volume, tPSA, and Q(max) were risk factors for predicting the development of AUR. CONCLUSION BPH patient's age, IPSS, Q(max), residual urine volume, prostate volume, transition zone volume, tPSA and fPSA, and PSA density all influence the occurrence of AUR, in which symptom severity, residual urine volume, total PSA and Q(max) are the principal risk factors for prognosing AUR.
Adult ; Age Factors ; Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; complications ; pathology ; Retrospective Studies ; Risk Factors ; Urinary Retention ; epidemiology ; etiology

OBJECTIVETo compare the effect of 5-fluorouracil (5-FU) portal vein infusion (PVI) for 7 days after radical resection, with intraluminal chemotherapy during operation for eliminating liver metastasis and elevating long-term prognosis in colorectal cancer.

METHODS162 colorectal cancer patients with radical resection were divided into portal vein chemotherapy group (group A, 82 cases) and intraluminal chemotherapy group (group B, 80 cases) randomly. In group A, 5-fluorouracil were infused with 1g per day constantly for 7 days after operation through portal vein catheters, which placed into greater omental vein and fixed on the abdominal wall. In group B, intraluminal chemotherapy was given and 5-fluorouracil 0.5 g was injected into the greater omental vein during operation.

RESULTSThe short-term complications and long-term effect in the two groups were compared by statistical software SPSS 8.0. Group A had more operative complications, and no statistical differences was found in hospital time and survival rate of the two groups. The 5-year survival rate is 76.7% (group A: 74.3%, group B: 79.2%), and the liver metastasis rate is 19.8%. There is no significant difference between the two group-survival curves. Multiple variable analysis suggested that Dukes' stage was the prognosis factor (P < 0.05).

CONCLUSIONSThe present study demonstrated that the two chemotherapy methods play an important role in preventing liver metastasis and improving the survival rate, and the intraluminal chemotherapy would be easier and simpler. The result should be further improved by using combined chemotherapy.

Adult ; Aged ; Antimetabolites, Antineoplastic ; administration & dosage ; Chemotherapy, Adjuvant ; Chemotherapy, Cancer, Regional Perfusion ; methods ; Colorectal Neoplasms ; drug therapy ; mortality ; therapy ; Combined Modality Therapy ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Portal Vein ; Survival Rate ; Treatment Outcome

OBJECTIVETo construct recombinant baculoviruses co-expressing three structural genes vp2, vp6 and vp7 of rotavirus, and assemble rotavirus-like particles (VLPs) in BmN cells.

METHODSHuman group A rotavirus was cultivated in MA104 cells, and the RNA was extracted and the three genes were obtained by RT-PCR. The PCR products were inserted into the transfer vectors pFBDM and pUCDM, respectively. A enhanced green fluorescent protein gene (egfp) driven by IE1 promoter was introduced into pFBDM to investigate the efficiency of infection. The expression baculoviruse was constructed by Tn7 and Cre-LoxP recombinant and transfected into BmN cells. The gene expression was determined by detecting 6-His tag fused into VP7 C-terminus, and the assembled VLPs were observed by transmission electron micrography.

RESULTSThree genes of rotavirus were cloned and BmMultiBac was constructed. The genes were expressed and the rotavirus-like particles assembled in BmN cells successfully as verified by ELISA and electron microscope.

CONCLUSIONWe have successfully constructed the recombinant baculovirus co-expressing the 3 structural genes of rotavirus, which provide the basis for producing protein complex containing multiple subunits and investigation of the structure of the macromolecules.

Animals ; Baculoviridae ; genetics ; metabolism ; Bombyx ; virology ; Capsid Proteins ; genetics ; Cell Line ; Gene Expression ; Genetic Vectors ; Rotavirus ; genetics ; metabolism

OBJECTIVEThis study aimed to investigate the effects of gene polymorphism of heat shock protein 70-2 (HSP 70-2) 1267A/G on the mRNA level HSP 70-2 mRNA and the protein level HSP 70 in human lung cancer.

METHODSForty six lung cancer patients diagnosed histopathologically between February and August 2008 from a hospital in zhengzhou were enrolled as the subjects in this study. Gene polymorphism of HSP 70-2 1276A/G in 46 patients with lung cancer was detected by PCR-RFLP. The mRNA levels of HSP 70-2 mRNA and the protein levels of HSP 70 in lung tissue and para-cancerous tissues of these subjects were determined by RT-PCR and Western blotting respectively.

RESULTSThe expression levels of HSP 70-2 mRNA (1.02 ± 0.30) and HSP 70 protein (0.44 ± 0.12) in the lung cancer tissues was significantly higher than that in para-cancerous tissues (0.19 ± 0.04, 0.12 ± 0.02). The relative levels of HSP 70-2 mRNA in the subjects with AA genotype (1.32 ± 0.22) were significantly higher than the patients with AG genotype or GG genotype (0.95 ± 0.17, 0.70 ± 0.16) at the site of 1267 (A/G) (P < 0.01); however, the relative protein levels of HSP 70 were 0.47 ± 0.13 (AA genotype), 0.42 ± 0.11 (AG genotype), 0.45 ± 0.11 (GG genotype), respectively, which showed no statistically significant difference (P > 0.05).

CONCLUSIONThe polymorphism of HSP 70-2 1267 (A/G) is highly associated with the transcription level of HSP 70-2 mRNA, but not with the expression level of HSP 70 protein.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Female ; Genotype ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Lung ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Neoplasm Staging ; Polymorphism, Single Nucleotide ; RNA, Messenger ; genetics

The Fusion (F) and Haemagglutinin-Neuraminidase (HN) genes of Newcastle disease virus (NDV) and the glycoprotein B (gB) gene of infectious laryngothracheitis virus (ILTV) as well as a LacZ reporter gene were all inserted into a nonessential gene of fowlpox virus (FPV) 017 strain by homologous recombination. The NDV and ILTV genes were each under the control of a fowlpox virus immediate early/late promoter (LP2EP2) while the LacZ reporter gene expression cassette was regulated by a P11 late promoter. A recombinant FPV harboring the F, HN and gB genes as well as the LacZ gene, designated as rFPV-F/HN/gB/LacZ, was obtained after ten cycles of blue plaque purification. The presence of the NDV and ILTV genes was confirmed by PCR. The expression of the recombinant proteins in rFPV-F/HN/gB/LacZ were characterized by Western blot (F and gB proteins) and indirect immunofluorescence test (F, HN and gB proteins). The results demonstrated that all four foreign proteins, which were encoded within a 10 kb gene fragment, could be expressed authentically and efficiently. Compared to the parental virus, rFPV-F/HN/gB/LacZ showed no obvious difference with respect to virus replication and cytopathogenic effects in chicken embryo fibroblasts (CEF) cell culture. Overall, our work suggests that FPV can be a useful live virus vector for the expression of multi- foreign genes against multiple avian pathogens.
Animals ; Cloning, Molecular ; Fibroblasts ; virology ; Fowlpox virus ; genetics ; Gene Expression ; Genetic Engineering ; methods ; HN Protein ; genetics ; Herpesvirus 1, Gallid ; genetics ; physiology ; Newcastle disease virus ; genetics ; physiology ; Plasmids ; genetics ; Transfection ; Viral Envelope Proteins ; genetics ; Viral Fusion Proteins ; genetics