Objective To investigate the therapeutic effect of fluvastatin in bleomycin-induced pulmonary fibrosis in rats. Methods Pulmonary fibrosis was induced in SD rats by intratracheal instillation of bleomycin A_ 5 . The rats were than divided randomly into three groups: the rats in the first group received daily fluvastatin 20mg/kg (group Flu),those of the second group received normal saline (group BLM) orally only,and those of controls (group N) received normal saline both intratracheally and orally. Five rats in each group were sacrificed 1,3,7,14 and 28 days after intratracheal instillation of bleomycin. Pathological changes in the lungs were evaluated by HE stain and Masson′s trichrome stain. Collagen content of the lung tissue was assessed by hydroxyproline concentration. Alveolar macrophages,polymorphonuclear leukocytes and lymphocytes in bronchoalveolar lavage fluid (BALF) were counted. Hyaluronic acid (HA) and laminin (LN) in BALF were determined by radioimmunoassay. Results The degree of alveolitis and pulmonary fibrosis in group Flu was improved as compared with that of group BLM. Hydroxyproline concentrations of group Flu were significantly lower than that of group BLM 7 days after bleomycin A_ 5 instillation. Total cell counts and percentage of polymorphonuclear leukocytes and lymphocytes in BALF were significantly reduced in group Flu. HA and LN levels in BALF were also lower in group Flu compared with group BLM. Conclusion Fluvastatin could alleviate bleomycin-induced pulmonary fibrosis in rats.

Objective:To observe the inhibitory effect of fluvastatin (Flu) on the proliferation of the rat lung fibroblasts cultured in vitro . Methods: Normal rat lung derived fibroblasts were cultured in media containing Flu. The influences of Flu on the growth curve of the fibroblasts were observed by cell count and MTT colorimetry. The inhibition effect of Flu serial dilutions on the fibroblasts proliferation was investigated. Influence of Flu on division index of the fibroblasts was analyzed by direct cell count. Chemical colorimetry was used to detect the hydroxyproline in the media. Results: Flu inhibited the proliferation of the normal rat lung fibroblasts in the manner of dose dependence( P

Sugarcane molasses containing large amounts of sucrose is an economical substrate for succinic acid production. However, Escherichia coli AFP111 cannot metabolize sucrose although it is a promising candidate for succinic acid production. To achieve sucrose utilizing ability, we cloned and expressed cscBKA genes encoding sucrose permease, fructokinase and invertase of non-PTS sucrose-utilization system from E. coli W in E. coli AFP111 to generate a recombinant strain AFP111/pMD19T-cscBKA. After 72 h of anaerobic fermentation of the recombinant in serum bottles, 20 g/L sucrose was consumed and 12 g/L succinic acid was produced. During dual-phase fermentation comprised of initial aerobic growth phase followed by anaerobic fermentation phase, the concentration of succinic acid from sucrose and sugarcane molasses was 34 g/L and 30 g/L, respectively, at 30 h of anaerobic phase in a 3 L fermentor. The results show that the introduction of non-PTS sucrose-utilization system has sucrose-metabolizing capability for cell growth and succinic acid production, and can use cheap sugarcane molasses to produce succinic acid.
Bioreactors ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; Fermentation ; Membrane Transport Proteins ; genetics ; Metabolic Engineering ; Molasses ; Saccharum ; chemistry ; Succinic Acid ; chemistry ; Sucrose ; chemistry

BACKGROUND: The pathological characteristics of pulmonary interstitial fibrosis are the proliferation of a large number of fibroblasts and the increasing deposition of matrix collagen that takes the place of normal lung structure. Fluvastatin can inhibit the proliferation of fibroblasts and many other cells.OBJECTIVE: To investigate the effects of fluvastatin in inhibiting the proliferation of rat lung fibroblasts cultured in vitro and its influence on bleomycin-induced pulmonary fibrosis and ventilation function.DESIGN: A randomized controlled trial.SETTING: Department of Respiratory Diseases, Xijing Hospital, Fourth Military Medical University of Chinese PLA; Teaching and Research Section of Pathology, Department of Basic Medicine, Fourth Military Medical University of Chinese PLA; Research Institute ofOrthopedics, Xijing Hospital,Fourth Military Medical University of Chinese PLA.PARTICIPANTS: The study was conducted in the laboratory of Department of Respiratory Diseases, Xijing Hospital of Fourth Military Medical University of Chinese PLA from January to December 2001. Thirty-one healthy adult male Sprague-Dawley(SD) rats of grade Ⅰ were selected in this study.INTERVENTIONS: The fibroblasts derived from the lung normal of one rat were cultured in vitro in media containing fluvastatin. The effect of fluvastatin on the growth curve and the effect of its different concentrations(0, 1 × 10-7,1 ×10-6, 1 ×10-5, 1 ×10-4, 1 ×10 3and 1 ×10-2 mol/L, fluvastatin of 0 mol/L was taken as the blank control group) in inhibiting the cultured cells were observed with MTT colorimetry. The effect of fluvastatin on the division index of the fibroblasts was analyzed by direct cell counting Hydroxyproline colorimetry was used to detect the influence of fluvastatin on the collagen secretion in the media. The other 30 SD rats were divided into six groups: normal control group, bleomycin-induced group and fluvastatin-treated groups(TH 1,TE1, TH15 and TL15 groups) named according to the date of giving fluvastatin,i. e. the 1st day and the 15th day, after the rats were given bleomycin A5. All the rats were killed 28 days later. The number of fibroblasts, the thickness of alveolar wall and the area of mesenchyma in lung tissue were measured by HE staining. The extracellular matrix and collagen in lung tissue were observed by Masson and sirius red staining, and hydroxyproline in lung tissue homogenates was measured.MAIN OUTCOME MEASURES: Fibroblast growth curve and division index of rat lung, hydroxyproline in the media and lung tissue homogenates,number of fibroblasts and the thickness of alveolar wall, the area of mesenchyma, extracellular matrix and collagen contents in lung tissue.RESULTS: Fluvastatin could inhibit the proliferation of the rat lung fibroblasts cultured in vitro(t=4.20 to 17.52, P < 0.01), and its inhibitory effect was increased with the increased dose of fluvastatin, which showed a dose-dependent effect. The 1 × 10-4 mol/L fluvastatin could completely inhibit the proliferation of the cultured cells, and the A490 value from the 2nd day on the fibroblasts by MTT colorimetry was not insignificantly different from those on the 1st day( P > 0.05) . The division index of the fibroblasts and secretion of collagen were obviously decreased by fluvastatin( t = 8. 037,P <0.01; t =3.99 to 10. 84, P <0.05 or P <0.01). In vivo, the number of fibroblasts, the thickness of lung alveolar wall, the area of mesenchyma and the content of hydroxyproline in lung tissue were significantly higher in bleomycin group than in control group( t =4. 62 to 11.93, P < 0. 01), while those in the fluvastatin-treated groups were lower than those in bleomycin group in different degrees( t = 2.69 to 7.65, P < 0.05 to 0.01 ) . The distribution of extracellular matrix and types Ⅰ and Ⅲ collagen in lung tissue were obviously increased in bleomycin group as compared with that in control group, but decreased in different degrees in fluvastatin-treated groups.CONCLUSION: Fluvastatin can significantly inhibit the proliferation of rat lung fibroblasts in vitro, suggesting that it may be an effective drug for pulmonary fibrosis. Treatment at earlier stage is more effective than at advanced stage.

Objective To detect the expression of Beclin1 and Bcl2 in invasive pituitary adenomas and to explore the relationship of Beclin1 and Bci2 in invasive pituitary adenomas and the relativity between the 2 genes.Methods 61 specimens were classified into invasive group (32 cases) and non-invasive group (29 cases) according to the comprehensive evaluation of invasive pituitary adenomas.lmmunofluorescence analysis and RT-PCR were adopted respectively to detect the protein and mRNA expressions of Beclinl and Bcl2.The difference and relativity of Beclin1 and Bcl2 expression in invasive group and non-invasive group were analyzed.Results 32 specimens of pituitary adenoma were invasive and 29 were non-invasive.Beclin1 protein and mRNA expressions were lower in the invasive group than in the non-invasive group (P ＜0.01 ).Bcl2 protein and mRNA expressions were higher in the invasive group than in the non-invasive group (P ＜0.01 ).Pearson related analysis showed that Beclin1 mRNA expression was negtively correlated with Bcl2 mRNA expression in the invasive group ( r =-0.42,P =0.028 ).Conclusions Beclinl expression is decreased in invasive pituitary adenomas.The invasiveness of pituitary adenoma is closely related to the high expression of Bcl2 protein and mRNA,and the low expression of Beclin1 protein and mRNA.The inhibition of the autophagy may lead to the enhancement of the invasiveness of pituitary adenomas and that inhibition may come from the interaction of Beclin1 and Bcl2.

Objective To study the association between the expression of periostin and the occurrence and progression of knee osteoarthritis (OA) in synovial fluid.Methods The expression level of periostin in the synovial fluid of healthy people and patients with different stages of OA was tested.Furthermore,60 surgical-induced OA rat model were divided into two groups,the sham operation group had only implemented slit suture,and the OA model group had one side anterior cruciate ligament transected.The expression of periostin in intra-articular injection samples were analyzed at 1,2,4,8,12 week.Fluorescence molecular tomography (FMT) were performed after surgery at 4,8,12 week on the surgery knee.Gross morphologic lesions on the tibial plateau in rats were visualized by India ink staining,toluidine blue staining,cartilage permeation test.The synovium were visualized by HE staining and periostin were detected by immunohistoc hemistry.The measurement data were compared by one factor analysis of variance test.Results The expression of periostin in cartilage was lower in late-stage OA than the one from normal and early-stage OA (F=13.95,P＜0.01).The FMT showed that there was no obvious change in the extent of chronic inflammation in the sham operation group,and the chronic inflammatory degree of the OA model group gradually increased as time went on.Toluidine blue staining and cartilage permeation test showed that the cartilage degeneration in rat model of OA became more and more serious with time.There was no stastically significant difference of the periostin in control groutp at different time stage (F=0.67,P=0.53).The periostin in the intra-articular increased at first and then decreased with the development of OA (F=11.0,P＜0.05).HE staining of synovial tissue showed that the degree of synovial hyperplasia was consistent with the degree of degeneration of joints.With the extension of time,the expression of periostin in synovial tissue increased gradually.Conclusion The expression of periostin in human synovial fluid is low in normal knee joint,increases in early and middle stages,and decreases in late stage.The rat model indicates that the expression of periostin increases first and then decreases with the development of OA,but the expression in synovium increases gradually with the development of OA.The increased expression of periostin in synovial fluid may serve as an early diagnostic marker for OA and downregulation of the periostin may be a start marker for the late OA.

Objective We have summarized the clinical characteristics of inappropriate antidiuresis(SIAD).Methods We adopted retrospective analysis to analyze the clinical and lab data of 40 cases.Results The most common causes of SIAD were malignant tumor,lung disease,and central nervous system disease.The five major abnormal lab data were:hypochloraemia,hypouricemia,hyponitremia,hypocalcemia,and low hematocrit.Conclusion It is important to diagnose SIAD as soon as possible,and patient presented hyponatremia combined with hypouricemia must be suspected to have SIAD.

Objective:To observe the effect of hypoxia on the dedifferentiation process of chondrocytes in vitro and explore the role of collagen prolyl 4-hydroxylase (C-P4Hs).Methods:Chondrocytes were treated with COCl 2 for different concentrations, and selecting the optimal COCl 2 concentration for hypoxia inductionwas100 μmol/L, the mouse costal chondrocytes were divided into the normal oxygen group and the hypoxia group, and the indexes of the 3rd generation 0.5-72 h and the 1st, 3rd, 5th and 7th generation at 72 h were detected respectively. The proliferation rate was determined by CCK8 method and cell count, and the dynamic changes of HIF-1α, P4Hα1, P4Hα2 and Col II in each group were detected by RT-qPCR, IF and Western blot. Results:Costal chondrocytes were cultured under different concentration of COCl 2 for 48 h. When COCl 2>150 μmol/L, the proliferation rate ( P<0.05) was significantly decreased. Normal oxygen and hypoxia induced rib chondrocytes for 0-72 h, and RT-qPCR showed significant increases in P4Hα2 and Col II mRNA in hypoxia group ( P<0.05). IF showed that HIF-1α and P4Hα2 accumulated in the nucleus under hypoxia, and P4Hα2 gradually entered the cytoplasm from the nucleus. Westernblot analysis showed that HIF-1α and P4Hα2 protein expressions were significantly increased in hypoxia group ( P<0.05). The expression of Col II protein in hypoxia group ( P<0.05) increased at the induction stage. CCK8 and cell count results showed that the proliferation rate and cell number of each generation in the hypoxic group were significantly increased ( P<0.05), and there was still potential for proliferation when the cells were transferred to the 6-7 generation. RT-qPCR showed that hypoxia group each generation cells P4Hα2, Col II mRNΑ were significantly increased ( P<0.05). Westernblot results showed that HIF-1α, P4Hα2 and Col II protein expressions were increased in each generation of hypoxia group ( P<0.05). ConcIusion:Increased expression of P4Hα2 through hypoxia induced HIF-1α can accelerate post-translational modification of Col II in chondrocytes and increase synthesis and accumulation of Col II. P4Hα2 may be responsible for increased proliferation rate and delayed dedifferentiation of chondrocytes cultured in vitro under hypoxia condition.

Methanol has become an attractive substrate for the biomanufacturing industry due to its abundant supply and low cost. The biotransformation of methanol to value-added chemicals using microbial cell factories has the advantages of green process, mild conditions and diversified products. These advantages may expand the product chain based on methanol and alleviate the current problem of biomanufacturing, which is competing with people for food. Elucidating the pathways involving methanol oxidation, formaldehyde assimilation and dissimilation in different natural methylotrophs is essential for subsequent genetic engineering modification, and is more conducive to the construction of novel non-natural methylotrophs. This review discusses the current status of research on methanol metabolic pathways in methylotrophs, and presents recent advances and challenges in natural and synthetic methylotrophs and their applications in methanol bioconversion.
Humans ; Methanol/metabolism* ; Metabolic Engineering ; Metabolic Networks and Pathways ; Biotransformation