Immunosuppressant mycophenolic acid biosynthesis employs a new globin-like enzyme for prenyl side chain cleavage

Mycophenolic acid (MPA, ) and its derivatives are first-line immunosuppressants used in organ transplantation and for treating autoimmune diseases. Despite chemical synthetic achievements, the biosynthetic formation of a seven-carbon carboxylic acid pharmacophore side chain of , especially the processes involving the cleavage of the prenyl side chain between DHMP () and DMMPA (), remains unknown. In this work, we identified a membrane-bound prenyltransferase, PgMpaA, that transfers FPP to to yield FDHMP (). Compound undergoes the first cleavage step a new globin-like enzyme PgMpaB to form a cryptic intermediate . Heterologous expression of genes in demonstrates that the second cleavage step (from to ) of is a cluster-independent process . Our results, especially the discovery of the broad tolerance of substrates recognized by PgMpaB, set up a strategy for the formation of "pseudo-isopentenyl" natural products using fungal globin-like enzymes.

Progress in fungal polyketide biosynthesis / 生物工程学报

Fungal polyketides display complex structures and variously biological activities. Their biosynthetic pathways generally contain novel enzyme-catalyzed reactions. This review provides a summary of recent research advances in molecular mechanism of the biosynthesis of fungal polyketides including highly-reducing polyketide synthases (HR-PKSs), non-reducing polyketide synthases (NR-PKSs), as well as polyketide-nonribosomal peptide synthase (PKS-NRPSs) and reducingnon- reducing polyketide synthase (HR-NR PKSs) hybrids. The elucidation of biosynthetic mechanism of many fungal polyketides provides guidance on the discovery of new biosynthetic gene cluster of fungal polyketide natural products and compounds with novel structures as well as their analogue.

Clinical efficacy of transurethral resection of bladder tumor combined with gemcitabine intravesical instillation in the treatment of superficial bladder cancer / 中国基层医药

Objective@#To investigate the effect of transurethral resection of bladder tumor (TURBt) combined with gemcitabine intravesical instillation in the treatment of superficial bladder cancer.@*Methods@#From May 2011 to January 2016, 112 patients with superficial bladder cancer were treated in the Seventh Hospital of Ningbo.According to the digital table, the patients were randomly divided into two groups: A group was treated by TURBt, and B group was treated by TURBt combined with gemcitabine.The postoperative complications, postoperative recurrence and quality of life were compared between the two groups.@*Results@#The operation time, indwelling catheter time and hospitalization time between the two groups had no statistically significant differences (all P>0.05). The postoperative complications of the two groups were frequent urination, dysuria and hematuria.The incidence rate of complications of A group was 39.29%, which was significantly higher than that of B group (χ2=36.597, P<0.05). The recurrence rate of B group was significantly better than that of A group (χ2=8.617, P<0.05). After treatment, the quality of life of the two groups was significantly improved compared with that before treatment, and compared with that of A group, the psychology, physiology, independence, social environment, quality of life scores of B group increased more significantly (t=29.217, 25.446, 19.517, 24.339, 36.001, all P<0.05).@*Conclusion@#Transurethral resection of bladder tumor combined with gemcitabine intravesical instillation in the treatment of superficial bladder cancer can ensure the treatment effect, at the same time, it can effectively reduce the postoperative complications and the emergence of various risk, reduce postoperative recurrence and improve the quality of life of patients, it has great significance and is worthy of promotion.

Course reform and thinking of the experimental teaching of pharmaceutical technology based on the case teaching / 中华医学教育探索杂志

The experimental teaching of Pharmaceutical Technology plays a critical role in improving the students' knowledge conversion,application,and innovation abilities in pharmaceutical engineering specialty.According to the problems existing in experimental teaching process,exploration and practice on the case teaching method and the student-centered teaching mode are introduced in this paper.The implementation of the teaching reform can effectively combine pharmaceutical technology theory with practice,and is conducive to the professional development of pharmaceutical engineering in our university.

A cross-sectional study of mental disorders in Ximeng Wa Autonomous County of Yunnan Province / 中国心理卫生杂志

Objective:To describe the epidemiological characteristics of mental disorders in Ximeng Wa Au-tonomous County and explore psychosocial risk factors of mental disorders. Methods:Two thousand three hundred and eighty one residents aged 1 8 years and over were sampled using multistage sampling in Ximeng Wa Autono-mous County in 2013. All respondents were investigated by face-to-face interview. Mental disorders were screened out by using the General Health Questionnaire (GHQ)and diagnosed according to the International Classification of Diseases Checklist (ICD-10-Checklist)criteria. Results:Life time prevalence of any mental disorder was 19. 86%(419/2110). The lifetime prevalence rates of substance use disorder,anxiety disorder,insomnia,mood disorder, schizophrenia were 12. 99%(274/2110),6. 30%(133/2110),2. 94%(62/2110),2. 32%(49/2110)and 1. 00%(21/2110)respectively. Male (OR=0. 43),older age (35 -49 years,OR=1. 78;50 -64 years,OR=2. 59;≥65 years,OR=3. 5 1 ),unmarried and other marital status (OR=0. 3 1 ),non-Wa and non-Lahu ethnic groups (OR=0. 29)were associated with neurotic,stress-related and somatoform disorders. Male (OR=2. 41),older age (35 -49 years,OR=2. 29;50-64 years,OR=3. 20;≥65 years,OR=4. 58),non-farmer and non-self-employed occupation (OR=0. 41),and non-Wa and non-Lahu ethnic groups (OR=0. 32)were associated with psychoactive substance use disorder. Male (OR =0. 35 ) and order age (≥65 years,OR =3. 05 ) were associated with mood disorders. Conclusion:Lifetime prevalence of any mental disorder,substance use disorders and anxiety disorders are high in Ximeng Wa Autonomous County. Measures should be strengthened against prevalence of mental disorders in ethnic minority areas.

Epigenetic regulation of secondary metabolite biosynthesis in filamentous fungi: a review / 生物工程学报

Secondary metabolites of filamentous fungi are important sources of new drugs, and their biosynthetic processes are regulated by numerous factors. Recent studies indicate that many filamentous fungal secondary metabolites are regulated by epigenetic modifications, which not only affect the titers of secondary metabolites, but also activate the cryptic gene clusters. This review summarizes recent advances of epigenetic application in filamentous fungal secondary metabolite biosynthesis, especially the types of fungal epigenetic modification and epigenetic remodeling of the fungal secondary metabolism. The application of epigenetic theory in filamentous fungi is becoming a new strategy for fungal strain improvement and a powerful method to obtain novel natural products.

Novel methods and strategies for strain improvement / 生物工程学报

Improvement of the productivity of industrial strains is an important field in micro-biology, because wild-type strains isolated from nature usually produce only a low level of antibiotics. Although random screening and simple rational screening are still effective without using genomic information, they are always time- and labor-consuming. With the broad application of recombinant DNA technology, protoplast fusion and X-omics, novel methods and strategies such as metabolic engineering, genome shuffling, system biology and system biotechnology, ribosome engineering, epigenetic modification are being exploited for the industry microbiology. In this review, we will focus on the progress of these novel methods and strategies for strain improvement in recent years.

A high throughput screening model for P-gp functional inhibitors / 中国药理学通报

Aim To establish a cell model for high throughput screening and search for novel inhibitors of P-gp.Methods A fluorescent cell-based assay was developed for application in high throughput screening.A library of 480 traditional Chinese medicine samples was screened. Results The value of Z′ was 0.72. Screening of a library of 480 traditional Chinese medicine identified 2 positive samples.Conclusions This HTS model for screening P-gp inhibitors was stable and sensitive.The 2 positive samples had good P-gp inhibitory activities.

Cloning expression and toxic effect on the host bacterial viability of an outer envelope protein from the strong virulent leptospira lai in China / 中国病理生理杂志

AIM: Construction of an eukaryote- E. coli shuttle expressing recombinant plasmid which expresses OmpL1 envelope protein of pathogenic Leptospira, serovar Lai strain 017. METHODS: The OmpL 1 gene was amplified by PCR from the leptospiral genome. Then it was cut with restriction enzymes and ligated to the plasmid pBK-CMV. The correct recombinant plasmid was screened out with analysis of restriction enzymes and PCR. After inducing the E. coli baring recombinant plasmid with IPTG,the complete protein of the bacteria was extracted for SDS-PAGE. At the same time, OD600 of the host bacteria was examined at different time after inducing or uninducing with IPTG. RESULTS: Five strains E. coli containing proper recombinant plasmids were screened out. Four strains E. coli expressed a new protein with a weight of 37 kD among them. With the expression of the heterogenous protein,the OD600 of the host bacteria decreased. CONCLUSION: The shuttle expressing plasmid of the OmpL 1 gene of strong virulent Leptospira strain 017 was successfully constructed. Furthermore,the recombinant plasmid expressed the expected OmpL 1 fusion protein in E. coli and the expression of the heterogenous protein had toxic effect on the host bacteria. This work was important for the future research of OmpL1 protein which relates to the diagnosis,new vaccine preparing and the pathogenic mechanism of leptospirosis.

Molecular genetic study on 17?-hydroxylase deficiency / 中华内分泌代谢杂志

Objective To study the molecular genetic mechanism of a patient with 17? hydroxylase (CYP17) deficiency. Methods Genomic DNA were abstracted from the blood of the patient, her parents and healthy control. The 8 exons of CYP17 gene were amplified, using 5 pairs of designed primers, with polymerase chain reaction (PCR), and the 8 exons were sequenced by the dideoxy terminator method to determined the mutation sites. The corresponding exons of the parents of the patients were also amplified and sequenced to determine the zygosity of the patient and the source of the gene variances. Results The analysis revealed that the patient (46, XY) was a compound heterozygote carrying two different inherited mutations on CYP17 gene, one from mother containing a point mutation Arg 96 (C G G)→ Gln(C A G) and the other from father containing a nine base deletion (CACTCTTTC) at amino acid position 487～489 (Asp Ser Phe) near the carboxyl terminus of P450c17. Conclusion The CYP17 gene of the patient with 17? hydroxylase deficiency is a compound heterozygous mutation. The mutation changes the amino acid sequence of P450c17 enzyme, which in turn affected the enzymatic activity. Arg 96 is essential in P450c17 enzyme activity. Deletion of Asp 487 Ser 488 Phe 489 in exon 8 may be a prevalent mutation causing P450c17 deficiency in Southeast Asia.