In order to improve the production of agro-antibiotic 2-16,the producing strain(Streptomyces ahygroscopicus var.huangshanensis) was treated by protoplast regeneration,ultraviolet radiation,NTG mutagenesis and low energy C~(+) ion implantation.At last,a high-yield strain No.515 was obtained.The production of ~()No.515 was increased by 223.10%.By using Plackett-Burman design and Response Surface Analysis provided by SAS software,the cultivation condition of No.515 was optimized.The amount of agro-antibiotic 2-16 was increased by 38.53% when the strain No.515 was cultivated in the optimum medium instead of the initial one.

Human kerotinocyte growth factor (hKGF) gene amplified by PCR was inserted into the shuttle vector pAdTrack-CMV to get the recombinant plasmid pAdTrack-CMV-hKGF, which was linearized with Pme I and transferred into Escherichia coli BJ5183 containing the adenoviral bone plasmid pAdEasy-1 to obtain the recombinant adenoviral plasmid pAdEasy-hKGF. The recombinant adenoviral plasmid was then transfected into HEK-293 cell lines via Lipofectamine 2000 to package and amplify the recombinant adenovirus containing hKGF gene detected by PCR. The recombinant adenovirus produced could effectively infect HaCat cells. The result of Western blotting showed that HaCat cells infected with the recombinant adenovirus expressed and secreted hKGF protein.

ObjectiveTo investigate the clinical features and risk factors of patient with Wegener's granulomatosis complicated with pulmonary infection.Methods Patients with Wegener's granulomatosis admitted to our hospital in the past 11 years were retrospectively analyzed.Comparisons between groups were performed by t tests or Fisher test.ResultsPulmonary infection occurred in 27 cases with an incidence rate of 29％.Twenty-six percent of pulmonary infections occurred at the initial diagnosis,and 44％ occurred within 6 months,while 30％ occurred later than 6 months.The clinical manifestations of pulmonary infection were productive cough (89％),hemoptysis (63％),fever and fatigue (56％),chest pain and pactoralgia (33％).The most common causative pathogen were bacteria(59％ ),fungi(37％ ),and tubercle bacillus(37％ ).Sinus infection(P=0.01),hypoproteinemia(P=0.03),hypoimmunoglobulinemia (P=0.007),and methylprednisolone pulse therapy(P=0.002) were the risk factors for pulmonary infection.ConclusionThe occurrence of Wegener's granulomatosis complicated with pulmonary infection is high within 6 months.The most common clinical manifestation is productive cough.The most common causative pathogens are bacteria,tubercle bacillus and fungi.Sinus infection,hypoproteinemia,hypoimmunoglobulinemia,and methylprednisolone pulse therapy are risk factors of pulmonary infection.

Connexin-43 (Cx43) expression in prostate cancer (PCa) cells and the potency of gap junctional intercellular communication (GJIC) in the cells were investigated, with an attempt to elucidate the reason why the so-called "bystander effect" mediated by thymidine kinase (TK) suicide gene therapy on PCa cells is not of significance and to explore the role of GJIC in PCa carcinogenesis. mRNA and protein expression of Cx43 in a PCa cell line PC-3m was detected by reverse-transcription polymerase chain reaction (RT-PCR) and strapt-avidin-biotin-enzyme complex (SABC) immunohistochemical staining, and inherent GJIC of PC-3m cells was assayed by scrape-loading and dye transfer (SLDT) assay. The expression of Cx43 in human normal and malignant prostate tissues was determined by SABC immunohistochemistry as well. It was found that Cx43 mRNA and protein expression in PC-3m cells was slightly reduced as compared with positive controls and the location of Cx43 protein was aberrant in cytoplasm rather than on membrane. Assessment of paraffin sections demonstrated that the expression of Cx43 protein in PCa cells was abnormally located and markedly diminished as compared with normal prostatic epithelial ones, displaying a negative correlation to the pathological grade (chi2=4.025, P<0.05). Additionally, capacity of inherent GJIC in PC-3m cells was disrupted, which was semi-quantified as (+) or (-). It was indicated that both down-regulated expression of Cx43 mRNA and aberrant location of Cx43 protein participated in the mechanisms leading to deficient GJIC in PC-3m cells. Lack of efficient GJIC is a molecular event, which may contribute not only to limited extent of "bystander effect", but also to initiation and progression of prostatic neoplasm.

The treatment and signaling pathway regulation effects of kidney- tonifying traditional Chinese medicine on osteoporosis have been widely studied, but without a systematic summary currently. This review comprehensively collected and analyzed the traditional Chinese medicine on the treatment and signaling pathway regulation of osteoporosis in recent ten years, such as Epimedium, Drynariae Rhizoma, Cnidium, Eucommia, Psoralen and Dipsacus. Based on the existing findings, we concluded the following conclusions: (1) kidney-tonifying traditional Chinese medicine treats osteoporosis mainly through BMP-Smads, Wnt/β-catenin, MAPK, PI3K/AKT signaling pathway to promote osteoblast bone formation and through OPG/RANKL/RANK, estrogen, CTSK signaling pathway to inhibit osteo?clasts of bone resorption. ① Epimedium, Drynariae Rhizoma, Cnidium and Psoralen up-regulate the key proteins and genes of BMP-Smads and Wnt/β-catenin signaling pathways to promote bone formation.② Epimedium, Drynariae Rhizoma, Cnidium, Eucommia, Psoralen, Dipsacusinhibit the bone resorption by mediating the OPG/RANKL/RANK signaling pathway. (2) Kidney-tonifying traditional Chinese medicine prevent and treat osteoporosis through a variety of ways: Icariin, Naringin, Osthol, Psoralen can regulate BMP-Smads, Wnt/β-catenin signaling pathway to promote bone formation, but also activate OPG/RANKL/RANK, CTSK and other signaling pathway to inhibit bone resorption. (3) The crosstalk of the signaling pathways and the animal experiments of the traditional Chinese medicine on the prevention and treatment of osteoporosis as well as their multi-target mechanism and comprehensive regulation need further clarification.

OBJECTIVETo explore etiologic fraction (EF) and interaction of hepatitis B virus (HBV) infection and other risk factors for primary hepatocellular carcinoma (PHC) in Guangxi, China.

METHODSA hospital-based case-control study including 500 PHC patients and 500 nontumorous patients was carried out in Guangxi. EF and interactions of HBV infection and other risk factors for PHC were analyzed by crossover analysis and nonconditional multiple logistic regression.

RESULTSHBV infection, family history of PHC, diabetes mellitus, eating raw fish, heavy alcohol consumption, frequently used drug, low income, mental oppression and blood type B all were risk factors for PHC. With EFs of 0.725, 0.186, 0.119, 0.486, 0.385, 0.438, 0.277, 0.607, 0.299, respectively and with etiologic fractions attributable to interaction [EF(A xB)] of 0.736, 0.643, 0.849, 0.551, 0.592, 0.618, 0.902, 0.577; and indices of interaction of 0.743, 0.651, 0.853, 0.560, 0.600, 0.626, 0.907, 0.586, respectively.

CONCLUSIONMain risk factors for PHC might include HBV infection, family history of PHC, diabetes mellitus, eating raw fish, heavy alcohol consumption, frequently used drug, low income, mental oppression and blood type B. HBV infection with other risk factors might exert synergistic action on developing PHC and increase the risk of PHC.

Adolescent ; Adult ; Aged ; Carcinoma, Hepatocellular ; etiology ; China ; epidemiology ; Data Interpretation, Statistical ; Female ; Humans ; Liver Neoplasms ; etiology ; Male ; Middle Aged ; Risk Factors

OBJECTIVETo investigate the precise locations of the blood vessels and nerves surrounding the seminal vesicles (SV) in men and provide some anatomical evidence for SV-related minimally invasive surgery.

METHODSWe observed the courses and distribution of the blood vessels and nerves surrounding SVs and obtained the data for positioning the SV neuroplexes in 20 male pelvises.

RESULTSOne branch of the neuroplexes was distributed to the SVs bilaterally with the neurovascular bundles, (2.85 ± 0.18) cm from the median sulcus of the prostate (MSP), while another branch ran through the Denonvillier fascia behind the SV, (0.81 ± 0.06) cm from the MSP. The arterial SVs (ASV) originated from the inferior vesical artery and fell into 4 types, 55% going directly to the SVs as one branch, 15% running between the SV and the ampulla of the deferent duct as another branch, 25% downward as 2 branches to the SV and between the SV and the ampulla of the deferent duct respectively, and 5% as the other ASVs. The shortest distance from the ASV through the prostatic neuroplexus to the posterior SV was (1.08 ± 0.09) cm.

CONCLUSIONIn SV resection, neuroplexus injury can be reduced with a bilateral distance of < 2.85 cm and a posterior distance of < 0.81 cm from the MSP, and so can bleeding by vascular ligation between the SV and the ampulla of the deferent duct.

Biopsy ; Humans ; Male ; Prostate ; blood supply ; innervation ; Seminal Vesicles ; blood supply ; innervation ; Vas Deferens ; blood supply ; innervation

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.
Adenoviridae ; genetics ; Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Fibroblast Growth Factor 10 ; genetics ; metabolism ; Humans ; Keratinocytes ; cytology ; metabolism ; Proteomics ; methods ; RNA, Messenger ; metabolism ; Recombinant Proteins ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Transfection ; Up-Regulation ; Voltage-Dependent Anion Channel 2 ; biosynthesis ; metabolism

The killing effects of herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) approach by the addition of several commonly clinical chemotherapeutic agents on hormone refractory prostate cancer (HRPC) cells PC-3m were investigated. After transferring of the HSV-tk gene into PC-3m cells, mRNA and protein expression of HSV-tk was detected by reverse-transcript polymerase chain reaction (RT-PCR) and strept avidin-biotin complex (SABC) immunohistochemical method. The killing effect of GCV, cisplatin (CDDP), etoposide (VP-16), vincristine (VCR), methotrexate (MTX), 5-fluorouracil (5-Fu), and suramin on PC-3m cells was evaluated by morphological assessment analysis, trypan blue exclusion assay and MTT assay respectively. Additionally, the cooperative effect of HSV-tk/GCV system combined with the above agents on the target cancer cells was determined by MTT. Furthermore, apoptosis and necrosis induced by GCV plus 5-Fu or suramin was analyzed by flow cytometry (FCM). The results showed that that there was HSV-tk mRNA and protein expression in pDR2-tk plasmid transduced PC-3m cell. Combination of GCV with VP-16, VCR, 5-Fu or suramin led to an enhanced cellular killing effect, but with CDDP resulted in a reduced one and with MTX in an approximate one. FCM revealed that synergistic use of GCV and 5-fu or suramin resulted in a rather large proportion of apoptosis and necrosis with the apoptosis index being 36.38% and 35.51%, and the proportion of necrosis being 33.05% and 28.87%, respectively. In conclusion, HSV-tk/CGV approach by addition of certain clinical available chemotherapeutic drugs brings on statistically significant enhanced cell killing over single-agent treatment. Our results highlight the potential for such new combination therapies for future treatments of HRPC.

Carcinoma ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Pancreatic Neoplasms ; genetics ; Spectral Karyotyping ; methods