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.

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.

Objective Preparation of a stable and reliable atrioventricular blockage ( AVB) animal model is of great im-portance to anti-arrhythmic drugs and biological engineering research .The aim of the article was to establish a rat model of AVB in-duced by chemical ablation, providing an effective animal model for the development of new drugs . Methods 60 adult SD rats were randomly divided into 4 groups(n=15): normal saline (NS) group, Verapmil (Ver) group, chemical ablation group 1 and chemical ablation group 2.0.9%NaCl (5 mg/kg) was injected into caudal vein of each rat in NS group .Verapmil (5 mg/kg) was injected into the caudal vein in Ver group.Anhydrous ethanol(50μL)was injected into atrioventricular groove area in chemical abla-tion group 1.Anhydrous ethanol(50μL) was injected into atrioventricular node area in chemical ablation group 2.The electrocardio-grams of the rats were examined by electrophysiological recorder.HE staining and Connexin 43 (CX43) immunohistochemical tech-nique were applied to atrioventricular junctions of the rats . Results A rat model of AVB was successfully established .Compared with NS group([45 ±2.24]per field of view), there was a significant decrease in the CX43 expressions of chemical ablation group 1 and group 2 ([15.20 ±2.23]per field of view, [22.10 ±4.70]per field of view)(P<0.05).Concerning NS group and Ver group, myocyte tissues of the atrioventricular nodal region were detected to remain in order without obvious changes and the expres-sion of CX43 immunoreactive protein was obvious under light microscope.As to chemical ablation group 1 and group 2, myocyte tissues were in degenerative necrosis and the expression of CX43 im-munoreactive protein was in lighter staining.Compared with Ver group, the incidence rate(73.3%, 60.6%) and the mortality rate (33.3%, 26.7%)of third-degree AVB decreased remarkably(P<0.05). Conclusion Chemical ablation can be induced to estab-lish a stable and reliable rat model of AVB , providing an effective
animal model for the research and development of new anti-arrhythmic drugs.

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 ; immunology ; Cytokines ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Immune System ; drug effects ; Male ; NF-kappa B p50 Subunit ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Plants, Medicinal ; Prostatitis ; drug therapy ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; Th1-Th2 Balance

Antineoplastic Agents ; therapeutic use ; Arsenicals ; therapeutic use ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Leukemia, Promyelocytic, Acute ; drug therapy ; Male ; Oxides ; therapeutic use ; Remission Induction ; methods ; Survival Rate ; Treatment Outcome

The modern physicians have different views on acupoint combination and acupuncture-moxibustion prescription and confuse them in clinical practice. It is significant to clarify the conception, connotation and relationship between them so as to normalize the therapeutic program of acupuncture and moxibustion and promote the standardization of acupuncture and moxibustion. Through the collection of relevant literature and analysis on the differences in the understandings among physicians, the conception, connotation and relationship between acupoint combination and acupuncture-moxibustion prescription are summarized. It is viewed that the acupoint combination is based on TCM theory. Under the guide of acupoint selection, in combination of the characters of clinical practice and acupoint indications, two or more than two acupoints of the same function are combined to enhance the collaborative effects of acupoints so as to achieve specific efficacy and improve clinical efficacy. Regarding acupuncture-moxibustion prescription, on the basis of disorder and syndrome differentiation of patients, the concrete therapeutic program is put forward, including acupoint composition and therapeutic method. Acupoint combination is the basic element of acupuncture-moxibustion prescription. Acupuncture-moxibustion prescription is the specific application of acupoint combination.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Moxibustion

Humans ; Accidents, Traffic ; Automobile Driving ; Risk Factors

AIM: To purify the water-extract of Sinisa recipe with macoroporous resigns. METHODS: Screening the purification condition by means of the content of active principle, fingerprint chromatography, the yield of dry extracts,et al. of the purified product. RESULTS: The purified product by type HP20 macoroporous resign is in good agreement with the active fraction of Sinisa in the composition of specific principles. CONCLUSION: HP20 type macoroporos resign is considered as optimum for active fraction of Sinisa recipe in purifying efficience.

Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.
Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Glycyrrhiza uralensis ; genetics ; Intramolecular Transferases ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Proteins ; genetics ; Recombinant Proteins ; metabolism ; Saccharomyces cerevisiae ; metabolism

Objective:To investigate the expression of wild type estrogen receptor(wER)and the ex-on-5 deleted ER(variant ER,vER)in human hepatocellular carcinoma(HCC)samples,and thereafteranalyze the possibility of HCC treatment by endocrine therapy.Methods:The mRNA expressions of wERand vER were analysed from 28 cases of HCC by RT-PCR.The expression of ER at the protein level wasdetected by immunohistochemistry(IHC).Results:IHC results showed that 39.3% of the HCC speci-mens expressed ER.The mRNA of wER was detected in 89.3%(25/28)of the HCC specimens whilethat of vER was detected in 96.4%(27/28).Twenty four out of 28 HCC cases(85.7%)expressedboth wER and vER.One out of 28 patients(3.5%)expressed only wER whereas 3 patients out of 28(10.7%)expressed vER only.Conclusion:Ninety six percent(27/28)of the HCC patients expressedvER,which suggests that the expression of vER is an important event in the development of HCC.