1.Comparative studies on codon usage bias of Ganoderma lucidum based on analysis of genomic and transcriptomic data.
Xiaoxuan ZHU ; Yingjie ZHU ; Jingyuan SONG ; Chao SUN ; Shilin CHEN
Acta Pharmaceutica Sinica 2014;49(9):1340-5
Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.
2.Optimization of Recombinant E. coli High-density Fermentation for Expressing Human-like CollagenⅡ
Haiyan CHANG ; Daidi FAN ; Yane LUO ; Xiaoxuan MA ; Yu MI ; Chenhui ZHU ; Lei CHI
Microbiology 1992;0(06):-
To study and optimize the fermentation parameters for expressing human-like collagenⅡduring E. coli high-density fermentation. The effects of pH, temperature, dissolved oxygen and induction instant on the cell growth and human-like collagenⅡproduction were investigated to optimize the fermentation conditions. The results demonstrated that the following conditions were beneficial for cell growth and foreign gene expression, controlling pH in phase induction at 6.8 and initial pH at 6.5, maintaining fermentation temperature and dissolved oxygen concentration was controlled at 34?C and 20% respectively, and implementing induction at the later logarithmic growth phase. Under the optimized condition, the cell density and human-like collagenⅡyield could reach 88.4 g/L and 14.2 g/L, respectively.
3.Anaerobic Biohydrogen Production Bacteria Selection and Its Optimization of Biohydrogen Fermentation Process
Shanshan LI ; Daidi FAN ; Xiaoxuan MA ; Yane LUO ; Yu MI ; Pei MA ; Chenhui ZHU ; Junfeng HUI
Microbiology 2008;0(09):-
This research adopted silt as the sample,and the five highest hydrogen production performing strains contained in the sample were isolated. The strain whose hydrogen production was the highest was identified as Enterobacter cloacae by the analysis of 16S rDNA sequencing and comparison. It is showed by Plackett-Burman Experimental Design that only glucose,citric buffer and reducing agent had significant effects on hydrogen production by Enterobacter cloacae FML-C1. The path of steepest ascent was undertaken to approach the optimal response region of those three factors. Central Composite Design(CCD) and Response Surface Methodology(RSM) were employed to investigate the interaction of the variables and to ascertain the optimal values of the factors,which finally led to the maximum hydrogen production(VH2) . The theoretical optimal medium conditions were:glucose 21.5 g/L,citric buffer 13.6 mL/L,reducing agent10.0 mL/L. The five tentative tests matched this model well. The final VH2 was up to 2347.4 mL/L,which was 127.42% enhanced in comparison to the original. The result shows that PB experiment design and RSM analytical method work well in selecting factors which have significant influences on the hydrogen production and,moreover,achieve the ideal optimal result.
4.Interaction of anti-thrombotic and anti-inflammatory activities of commonly used traditional Chinese medicine for promoting blood circulation and removing blood stasis revealed by network pharmacology analysis.
Ming Lü ; Taiyi WANG ; Xiaoxuan TIAN ; Xinhui SHI ; Guanwei FAN ; Yan ZHANG ; Yan ZHU
Acta Pharmaceutica Sinica 2015;50(9):1135-41
Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.
5.Effects of electroacupuncture at points of a traditional acupuncture formula that induces labor with different parameters on uterine contractility in late-stage pregnant rats.
Yingru CHEN ; Hongwen YUAN ; Fuzheng SHU ; Xiaoxuan REN ; Liangxiao MA ; Renquan LIU ; Jiang ZHU
Journal of Integrative Medicine 2011;9(1):91-9
To select the most effective parameters of electroacupuncture (EA) at Hegu (LI4) and Sanyinjiao (SP6), a traditional acupuncture formula that induces labor, by comparing its effects on uterine contraction in late-stage pregnant rats, so as to improve the effects of acupuncture on induction of labor and its clinical maneuverability and provide the basis for further mechanism research.
6.Development of the devices for synthetic biology of triterpene saponins at an early stage: cloning and expression profiling of squalene epoxidase genes in panax notoginseng.
Yunyun NIU ; Xiaoxuan ZHU ; Hongmei LUO ; Chao SUN ; Linfang HUANG ; Shilin CHEN
Acta Pharmaceutica Sinica 2013;48(2):211-8
Synthetic biology of traditional Chinese medicine (TCM) is a new and developing subject based on the research of secondary metabolite biosynthesis for nature products. The early development of synthetic biology focused on the screening and modification of parts or devices, and establishment of standardized device libraries. Panax notoginseng (Burk.) F.H.Chen is one of the most famous medicinal plants in Panax species. Triterpene saponins have important pharmacological activities in P. notoginseng. Squalene epoxidase (SE) has been considered as a key rate-limiting enzyme in biosynthetic pathways of triterpene saponins and phytosterols. SE acts as one of necessary devices for biosynthesis of triterpene saponins and phytosterols in vitro via synthetic biology approach. Here we cloned two genes encoding squalene epoxidase (PnSE1 and PnSE2) and analyzed the predict amino acid sequences by bioinformatic analysis. Further, we detected the gene expression profiling in different organs and the expression level of SEs in leaves elicited by methyl jasmonate (MeJA) treatment in 4-year-old P notoginseng using real-time quantitative PCR (real-time PCR). The study will provide a foundation for discovery and modification of devices in previous research by TCM synthetic biology. PnSE1 and PnSE2 encoded predicted proteins of 537 and 545 amino acids, respectively. Two amino acid sequences predicted from PnSEs shared strong similarity (79%), but were highly divergent in N-terminal regions (the first 70 amino acids). The genes expression profiling detected by real-time PCR, PnSE1 mRNA abundantly accumulated in all organs, especially in flower. PnSE2 was only weakly expressed and preferentially in flower. MeJA treatment enhanced the accumulation of PnSEI mRNA expression level in leaves, while there is no obvious enhancement of PnSE2 in same condition. Results indicated that the gene expressions of PnSE1 and PnSE2 were differently transcribed in four organs, and two PnSEs differently responded to MeJA stimuli. It was strongly suggested that PnSEs play different roles in secondary metabolite biosynthesis in P. notoginseng. PnSE1 might be involved in triterpenoid biosynthesis and PnSE2 might be involved in phytosterol biosynthesis.
7.Clinic study of serum product advanced oxidation protein changes of patients with chronic obstructive pulmonary disease
Jilong SHEN ; Hongjun GU ; Xiaohong PEI ; Qingtai ZHOU ; Ling LI ; Xuemei ZHOU ; Xiaoxuan ZHU
Clinical Medicine of China 2014;30(11):1184-1187
Objective To investigate the serum levels of advanced oxidation protein products (AOPP) in patients with chronic obstructive pulmonary disease (COPD) as well as explore the relevance of its clinical significance AOPP and COPD.Methods Fifty-four patients with mild/moderate COPD (COPD group) were enrolled in this study,who were treated in the No.100th Hospital of the Chinese People's Liberation Army from Apr.2011 to Nov.2013.Thirty healthy volunteers (control group) at same period were selected as control group and the general condition of two groups were matched.AOPP,superoxide dismutase(SOD) and malondialdehyde (MDA) levels were measured.Results The serum AOPP,MDA and SOD in control group were (45.78 ± 12.54) μmol/L,(2.96 ± 0.55) μmol/L and (78.40 ± 8.37) kU/L respectively.And its were (68.93 ± 10.62) μmol/L,(6.07 ± 2.44) μmol/L and (53.66 ± 5.99) kU/L respectively in COPD group.The differences were statistically significant (t =-8.57,-9.14,14.38 ; All P values were less than 0.01).The serum AOPP,MDA and SOD in mild COPD group were (65.56 ±9.65) μmol/L,(4.21 ± 1.83) μmol/L and (62.97 ± 6.28) kU/L respectively,and (71.79 ± 11.37) μmol/L,(7.43 ± 3.12) μmol/L and (41.25 ± 5.89) kU/L respectively in moderate CODP group.The differences were statistically significant (t =-2.17,-4.80,13.00; P < 0.05 or P < 0.01).Conclusion The increased levels of serum AOPP is important pathological changes in patients with COPD,which may be involved in the occurrence and development of COPD,and is the reaction of oxidative stress injury in patients with COPD early sensitive indicator.
8.Prevalence and genotypes of Chlamydia psittaci in pigeons in Jilin Province, Northeastern China
Qiuxia YAO ; Xiaoxuan ZHANG ; Kai CHEN ; Jiangang MA ; Wenbing ZHENG ; Xiaoqin XU ; Xingquan ZHU
Chinese Journal of Zoonoses 2017;33(2):104-109
Chlamydia psittaci is a causative agent of psittacosis,which can infect a wide range of hosts including birds and humans.However,information regarding C.psittaci infection in pigeons is scarce.In the present study,a total of 399 fecal samples from pigeons were collected from Jilin Province,northeastern China,between March and May 2015,and examined by nested PCR amplification of outer membrane protein A (ompA) gene.The overall Chlamydiosis prevalence was 5.01% (21/399),with 3.19% in Changchun City and 9.40% in Jilin City.Furthermore,breed was the major risk factor associated with Chlamydia infection in pigeon,boiler pigeons had a prevalence of 7.49%,whereas no C.psittaci was detected in racing pigeons.Sequence analysis of the ompA gene revealed that all the identified isolates represented C.psittaci genotype B.Our results firstly indicated the presence of zoonotic C.psittaci in boiler pigeons in Jilin Province,northeastern China,and effective measures should be implemented to reduce the risk of C.psittaci transmission from pigeons to humans.
9.Screening of the effective cellulose-degradable strain and its application in the production of cellulose bioethanol
Pengfei GAO ; Daidi FAN ; Pei MA ; Yane LUO ; Xiaoxuan MA ; Chenhui ZHU ; Junfeng HUI
Journal of Pharmaceutical Analysis 2009;21(3):184-188
Strains from the cellulose-containing environment were collected. Primary screening(by filter-paper Hutchison solid culture medium and sodium carboxymethylcellulose solid culture medium) and reelection(by filter-paper inorganic salt culture medium and sodium carboxymethylcellulosc Congo red coltnre medium) indicated that five strains obtained were best suited for high performance cellulose degradation. Determination of sodium carboxymethylcellulose activity(CMCA) and filter paper activity(FPA) was accomplished for each of the five. The strongest of the five in CMCA and FPA was applied to the production of cellulose bioethanol by separate hydrolysis and fermentation(SHF) and simultaneous saccharification and fermentation(SSF) respectively.
10.A multicenter clinical study of 280 cases of staphylococcal peritoneal dialysis-associated peritonitis
Xinyang LI ; Liming YANG ; Xueyan ZHU ; Xiaoxuan ZHANG ; Jing ZHAO ; Shichen LIU ; Xiaohua ZHUANG ; Yanfeng WU ; Ping LUO ; Wenpeng CUI
Chinese Journal of Nephrology 2021;37(4):321-326
Objective:To investigate the incidence, drug sensitivity and drug resistance characteristies, and theraputic effect of staphylococcal peritoneal dialysis-associated peritonitis (PDAP), aim to provide clinical evidences for standardizing treatment therapy of staphylococcal PDAP. Methods:Clinical data of PDAP patients admitted to the Second Hospital of Jilin University, the First Hospital of Jilin University-the Eastern Division, Jilin Central Hospital and Jilin First Automobile Work General Hospital during January 1, 2013 and December 31, 2019 were retrospectively collected. The results of etiology, drug sensitivity and drug resistance of staphylococcal PDAP patients were collected. According to the pathogenic bacteria, patients were divided into staphylococcus aureus group ( n=48) and coagulase-negative staphylococcus group ( n=232). According to the results of methicillin resistance, patients were divided into drug-resistant group ( n=71) and drug-sensitive group ( n=30). The prognosis of antibiotic therapy in each group were compared. Poisson regression was used to test the changing trend of the incidence of staphylococcal PDAP. The changes of drug sensitivity and drug resistance of staphylococcus were compared between 2013 and 2019 by linear trend χ2 test. Results:A total of 1 085 cases of PDAP occurred in 625 patients were screened, and 280 cases of staphylococcal PDAP were finally included. The incidences of staphylococcal PDAP, staphylococcus aureus PDAP and coagulase-negative staphylococcal PDAP were 0.063 times per patient year, 0.010 times per patient year and 0.053 times per patient year respectively. In addition, the incidence of PDAP caused by staphylococcus, staphylococcus aureus and coagulase-negative staphylococcus decreased year by year (all P<0.05). With the change of years, the sensitivity rate of staphylococcus to rifampicin increased, while the sensitivity rate of staphylococcus to moxifloxacin decreased (both P<0.05). The drug resistance rate of staphylococcus to levofloxacin increased ( P<0.05). The staphylococcus aureus group was more prone to refractory PDAP and catheter removal than that in coagulase-negative staphylococcus group, and the recurrence rate was higher than that in coagulase-negative staphylococcus group (all P<0.05). The proportion of vancomycin used during the whole course of antibiotic therapy in drug-resistant group was higher than that in drug-sensitive group ( P<0.05). Conclusions:The incidence of staphylococcal PDAP decreases year by year, and the drug sensitivity characteristics of staphylococcus also change. The therapeutic outcomes of staphylococcus aureus PDAP are worse than that of coagulase-negative staphylococcus.