2.Study of Breeding Saccharomyces cerevisiae with Improved Temperature and Ethanol Tolerance by Genome Shuffling
Hao WANG ; Hang WANG ; Chun MENG ; Yang-Hao GUO ;
Microbiology 1992;0(04):-
By UV induced mutagenesis of protoplasts of Saccharomyces cerevisiae strain f4, f5 and f6, and screening on plates containing different concertration of ethanol at different temperature, we obtained improved strains, such as f4.2, f5.1, f6.2, f4.5. By using a DES to deal with all the improved strains, We obtained two mutants, f5.1.1 and f4.2.1,which have improved ethanol torlerance. We made use of genome shuffling to generate improved strains in this work. We shuffled twice these improved strains by protoplast fusion and finally obtained strains with higher temperature and ethanol tolerance. we also identified shuffled strains that produced more ethanol by shake-flask experments. At the 35℃, the ethanol yield of R24 strain got to 12.93% (W/V), and were almost 5% higher than that of stain f4.
3.Silencing of miR-21 influences the function of colon cancer cell line HT-29 and the expression of PDCD4
Yupeng REN ; Chun SONG ; Hao ZHANG
China Oncology 2015;(1):6-12
Background and purpose: PDCD4 may be inhibited by miR-21 to regulate the malignant behaviors of colon cancer such as invasion and migration. This study aimed to explore the function of colon cancer HT-29 cell lines by downregulating miR-21 expression and discuss the mechanisms and relationship between miR-21 and PDCD4 in colon cancer malignant behaviors. Methods:simiR-21 was transfected into colon cancer cell line HT-29 to downregulate the expression of miR-21. Proliferation, apoptosis, migration and invasion were detected by MTT, flow cytometry and Transwell assay after transfection. PDCD4 expression was detected by Western blot and qRT-PCR. Results:The qRT-PCR analysis result proved that the transfection efifciency was 60%-65%. MTT analysis result showed that the proliferations of HT-29 cells were inhibited after the transfection of miR-21 for 72, 96, 120 h (t=1.276, P<0.05;t=3.276, P<0.01;t=4.523, P<0.01). Comparing with si-negative control and miR-21 groups, lfow cytometry result showed that the apoptosis rate was increased after miR-21 expression downregulated (t=2.132, P<0.05;t=3.524, P<0.05). Transwell assay result showed that migration (t=2.423, P<0.05; t=3.153, P<0.05) and invasion(t=3.245, P<0.05; t=5.236, P<0.05) were inhibited;Western blot result showed that PDCD4 expression was up-regulated at protein level(t=2.342, P<0.05;t=4.215, P<0.05);qRT-PCR result showed that PDCD4 expression was up-regulated at mRNA level(t=2.261, P<0.05; t=3.492, P<0.05). Conclusion: The proliferation, migration and invasion are the inhibited, and apoptosis is attenuated after miR-21 downregulated by simiR-21 transfection, PDCD4 expression is up-regulated. miR-21 may enhance the malignant behavior of cancer cells by downregulating the PDCD4 expression, miR-21 might be a target gene for colon cancer therapy.
5.Clinical analysis of 109 cases of isolated fetal pyelectasis and pregnancy outcome
Hao ZHU ; Chun SHEN ; Xiaotian LI
Chinese Journal of Perinatal Medicine 2015;18(9):683-686
Objective To investigate the prognosis of isolated fetal pyelectasis detected by ultrasound.Methods A total of 109 cases of isolated fetal pyelectasis (renal pelvis anteroposterior diameter ≥ 5 mm by ultrasound screening at any gestational age without structural or chromosome abnormalities) detected by prenatal ultrasound screening from March 2004 to July 2014 in Obstetrics & Gynecology Hospital of Fudan University,were delivered and followed up until neonatal period on chromosome examination,prenatal B ultrasound and outcome of neonates.Receiver operating characteristics curves were plotted and used to predict the optimal critical point of poor prognosis and the warning point of follow-up.Results Among the 109 cases,83 cases tended to have a natural recovery during pregnancy,71 of them had normal renal pelvis.Two of the neonates died,9 cases needed surgical treatment,and 98 cases had normal renal pelvis or need follow-up only.The area under the receiver operating characteristics curves was 0.860 (95%CI:0.860± 1.96 × 0.112).The optimal critical point of poor prognosis was determined at ≥ 11 mm in anteroposterior diameter with sensitivity 81.8%,and specificity 83.7%,and the warning point of prenatal follow-up was at ≥ 7 mm in anteroposterior diameter with sensitivity 100.0% and specificity 50.0%.Conclusions The prognosis of isolated fetal pyelectasis is mostly good.The fetus with pyelectasis thicker than 7 mm should be followed-up closely during prenatal and neonatal period,and the fetus with pyelectasis thicker than 11 mm is likely to have poor prognosis in neonatal period.
7.MeaSurement of Relative Affinity and Specificity of Monoclonal Antibodies
Chun-Hai HAO ; Yuan-Yao CHEN ;
Chinese Journal of Immunology 1985;0(05):-
To measure the relative affinity and type Specificity of neutralizing monoclonal antibodies against the hexon antigen of adenovirus type 7,a simple ELISA double antibody binding system has been developed.By this method,ralative affinity of nine MAbs was estimated from the antibody concentrations at approximately 50% of plateau binding and were ranked.The theoretical basis for this method was discussed. Hybridomas secreting antibodies of desired affinity can be selected at an early stage after fussion by measuring relative affinity of hybridomas supernatants.The type specificity was judged by the difference between end-point concentration against adeno- virus type 7 and type 3.And the relation between affinity and specificity of MAbs was discussed.
8.Effect of total flavones of Epimedium leptorrhizum on osteoporosis in ovariectomized rats.
Chun-Yu CAO ; Ting LIU ; Lian-Qiang HUI ; Ran HAO
China Journal of Chinese Materia Medica 2014;39(10):1913-1917
OBJECTIVETo observe the effect of total flavones of Epimedium leptorrhizum (YYH-C) on osteoporosis in ovariectomized rats.
METHODOvariectomized female rats were randomly divided into the model group, YYH-C lower, middle and high dose (0.7, 1.4, 2.8 g x kg(-1)) groups, the positive drug Bujiale (0.15 mg x kg(-1)) group, and the sham group. The rats were orally ad-ministrated with drugs for three months. Parathyroid hormone (PTH), procollagen I N-terminal peptide (PINP), alkaline phosphatase (ALP), calcium (Ca) and phosphrous (P) in serum were detected. Femur bones and vertebrae bones of left side were collected to determined bone metrological indexes, including bone mineral density (BMD), bone Ca, and bone ash weight/dry weight percentage. Femur bones of right side were collected to for a morphological observation of bone.
RESULTCompared with the sham group, the model group showed significantly higher PTH and ALP content but obviously lower PINP and Ca content. The three YYH-C 3 groups could resist the decrease of PINP. Specifically, low and middle dose groups could remarkably inhibit the increase of PTH, and the high dose group could increase the Ca content in serum, but without significant effect on the rise in ALP. There was no significant difference in P content in serum in each group. BMD, ash weight/dry weight percentage, Ca and P content of the model group were significantly lower than those in the sham group. The high dose YYH-C group could significantly increase BMD. All of the three YYH-C groups could notably increase ash weight/dry weight percentage and Ca, P content in femur bones and vertebrae bones. YYH-C could significantly increase average thickness, area, area percentage of bony trabeculae, cortical bone area percentage of femoral shaft and the number of osteoblasts on the surface of bony trabeculae, and decrease the number of osteoclasts.
CONCLUSIONYYH-C can effectively control the bone mass loss of rats with ovariectomy-induced osteoporosis, prevent the changes in bone microstructure, and inhibit bone absorption, so as to resist high turn-over osteoporosis after ovariectomy. [Key words] total flavones of Epimedium leptorrhizum; ovariectomized rat; osteoporosis
Alkaline Phosphatase ; metabolism ; Animals ; Bone Density ; drug effects ; Calcium ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Epimedium ; chemistry ; Female ; Flavones ; administration & dosage ; Humans ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; Parathyroid Hormone ; metabolism ; Rats ; Rats, Sprague-Dawley
9.Immune mechanisms of the active ingredients of Chinese medicinal herbs for chronic prostatitis.
Hao WANG ; Yu-chun ZHOU ; Jian-guo XUE
National Journal of Andrology 2016;22(1):63-66
Chronic prostatitis is a common male disease, and its pathogenesis is not yet clear. Most scholars believe that oxidative stress and immune imbalance are the keys to the occurrence and progression of chronic prostatitis. Currently immunotherapy of chronic prostatitis remains in the exploratory stage. This article relates the active ingredients of 5 Chinese medicinal herbs (total glucosides of paeony, tripterigium wilfordii polglycosidium, curcumin, geniposide, and quercetin) for the treatment of chronic prostatitis and their possible action mechanisms as follows: 1) inhibiting the immune response and activation and proliferation of T-cells, and adjusting the proportion of Th1/Th2 cells; 2) upregulating the expression of Treg and enhancing the patient's tolerability; 3) suppressing the activation of the NF-kB factor, reducing the release of iNOS, and further decreasing the release of NO, IL-2 and other inflammatory cytokines, which contribute to the suppression of the immune response; 4) inhibiting the production of such chemokines as MCP-1 and MIP-1α in order to reduce their induction of inflammatory response. Studies on the immune mechanisms of Chinese medicinal herbs in the treatment of chronic prostatitis are clinically valuable for the development of new drugs for this disease.
Chemokines
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immunology
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Cytokines
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immunology
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Immune System
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drug effects
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Male
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NF-kappa B p50 Subunit
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metabolism
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Nitric Oxide Synthase Type II
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metabolism
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Plants, Medicinal
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Prostatitis
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drug therapy
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immunology
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T-Lymphocytes, Regulatory
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drug effects
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Th1-Th2 Balance
10.Effects of SPARC on expression of TGF-βl and collagen type Ⅰ in keloid fibroblasts
Chun WANG ; Xiaojing LI ; Chaohua LIU ; Xiaojuan WENG ; Dongyue HAO
Chinese Journal of Medical Aesthetics and Cosmetology 2011;17(5):377-380
Objective To observe the effects of secreted protein,acidic and rich in cysteine (SPARC) on the expression of TGF-β1 and collagen type Ⅰ in cultured human keloid fibroblasts by real-time fluorescence quantitative RT-PCR.Methods In vitro keloid fibroblasts were stimulated by different concentrations of recombinant human SPARC,and with the control group for comparison,real-time fluorescence quantitative RT-PCR to detect expression of TGFβ1 and collagen type Ⅰ.Results Compared with the control group,the expression of TGF-β1 and collagen type Ⅰ was significantly increased in the experimental group.Conclusions SPARC could enhance the expression of TGF-β1 and collagen type Ⅰ in keloid fibroblasts significantly.