1.Effects of chemically modified sugarcane bagasse on butanol production by immobilized Clostridium acetobutylicum XY16.
Xiangping KONG ; Aiyong HE ; Jianan CHEN ; Wufang CHEN ; Chunyan YIN ; Pan CHEN ; Hao WU ; Min JIANG
Chinese Journal of Biotechnology 2014;30(2):305-309
Sugarcane bagasse modified by polyethylenimine (PEI) and glutaraldehyde (GA) was used as a carrier to immobilize Clostridium acetobutylicum XY16 in the process of butanol production. The effects of chemically modified sugarcane bagasse on batch and repeat-batch fermentations were investigated. Batch fermentation was conducted with an addition of 10 g/L modified sugarcane bagasse and 60 g/L glucose, resulting in a high solvent concentration of 21.67 g/L and productivity of 0.60 g/(L x h) with the treatment of 4 g/L PEI and 1 g/L GA. Compared to the fermentations by free cells and immobilized cells on unmodified sugarcane bagasse, the productivity increased 130.8% and 66.7%, respectively. The fibrous-bed bioreactor also maintained a stable butanol production during repeat-batch fermentations, achieving a maximum productivity of 0.83 g/(L x h) with a high yield of 0.42 g/g.
Batch Cell Culture Techniques
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Bioreactors
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Butanols
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metabolism
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Cells, Immobilized
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Cellulose
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metabolism
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Clostridium acetobutylicum
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metabolism
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Fermentation
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Saccharum
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chemistry
2.Prokaryotic Expression and Potential Application of the Truncated PCV-2 Capsid Protein
Zhongzi LOU ; Zhiyong LI ; Gang WANG ; Jianqiang LI ; Xi LAN ; Xuerui LI ; Xiangping YIN ; Jixing LIU ; Sidang LIU
Virologica Sinica 2010;25(2):86-97
Three pairs of specific primers were designed to amplify the F2-1, F2-2 and XF2-2 truncated sequences of ORF2 which encodes the capsid protein of porcine circovirus type 2 (PCV-2). The F2-1 sequence had most of the NLS region of ORF2, but the F2-2 and XF2-2 genes had the NLS region deleted. Truncated genes were subcloned into pET-32a(+) vectors to construct recombinant fusion expression vectors. The vectors were then transformed into Rosetta(DE3) E. coli and expressed by induction of IPTG. Expressed proteins were detected by western blotting and ELISA. The protein with best immunoreactivity was confirmed and selected, then utilized to inoculate SPF rabbits to prepare polyclonal antibodies. The protein and prepared polyclonal antibody were utilized to detect sera samples against PCV-2 from Shandong province and PCV-2 particles in PK-15 cells. In our study, three recombinant fusion proteins were successfully obtained, and the molecular weights of fusion proteins were 35.9 kDa, 33.6 kDa and 38.6 kDa respectively detected by SDS-PAGE. All of the proteins showed positive reaction with anti-PCV-2 antisera, and His-XF2-2 showed better immunoreactivity than the others. The protein of His-XF2-2 was coated as antigen in ELISA to detect the seroprevalence of PCV-2 in certain districts of Shandong province, the seropositivity rate was 27.7 % (73/264). Specific fluorescence and positive signals for PCV-2 could be detected in PK-15 cells inoculated with PCV-2 with the participation of prepared antibodies against His-XF2-2 in IFA and IPMA. Experimental results indicated that the truncated PCV-2 ORF2 gene containing most of the NLS region was successfully expressed in E. coli, and His-XF2-2 was demonstrated to have better immunoreactivity with anti-PCV-2 antisera than the other two fusion proteins. His-XF2-2 and prepared polyclonal antibodies against it had a satisfactory capability in detecting PCV-2 infection.
3.Herbs for calming liver and suppressing yang in treatment of hyperthyroidism with hyperactive liver yang: herbal effects on lymphocyte protein expression.
Xiangping LI ; Tao YIN ; Guangwei ZHONG ; Wei LI ; Yanhong LUO ; Lingli XIANG ; Zhehao LIU
China Journal of Chinese Materia Medica 2011;36(14):1997-2004
OBJECTIVETo observe the herbal effects on hyperthyroidism patients with syndrome of hyperactivity of liver-Yang by method for calming the liver and suppressing Yang and investigate its effects on the lymphocyte protein expression. This approach may lay a foundation for the further investigation of the curative mechanisms of calming the liver and suppressing Yang treatment.
METHODA total of 48 hyperthyroidism patients with syndrome of hyperactivity of liver-Yang were randomly divided into treatment group and control group. The treatment group was treated by method for calming the liver and suppressing Yang in accordance with traditional Chinese medicine (TCM) and the control group with thiamazole tablets for three periods of treatment The therapeutic effects, the score of TCM symptom, electrocardiogram (P wave), thyroid hormones and ultrasound were observed in both groups before and after the treatment. The side effects in the treatment course were observed in both groups. The level of differential protein expression was analyzed by two-dimensional electrphoresis and matrix assisted laser desorption/ionizaton time-of-flight mass spectrometry.
RESULTThe treatment group has the effect on stepping down the heart rate, cutting down the P wave amplitude changes, regulating the level of thyroid hormones and decreasing the volume of thyromegaly. There are not statistically significant between the treatment group and control group. However, the treatment group has obviously better effect on regulating TCM symptom and decreasing the side reaction than the control group (P<0.05). There are not statistically significant on the total effective between the treatment group and control group. The average spots in lymphocyte for normal people, before and after treating hyperthyroidism patients with syndrome of hyperactivity of liver-Yang were (429 +/- 31), (452 +/- 28) and (437 +/- 36) spots respectively. Eight down-regulated protein expressions and 11 up-regulated protein expressions were obtained in the hyperthyroidism patients with syndrome of hyperactivity of liver-Yang and normal people. Five strengthened expressions of protein were also obtained in 8 down-regulated expressions of protein and 8 lower expressions of protein in 11 up-regulated expressions of protein before and after treating the migraine patients with syndrome of hyperactivity of liver-Yang. Ten of the total 8 differential protein spots were successfully identified by MALDI-TOF-MS. The functions of these proteins were involved in metabolism associated, transportation, antioxidation, sigal transduction and immume associated protein, etc. according to information provided by NCBI and MSDB database.
CONCLUSIONIn this study, the TCM complex prescription with herbs for calming the liver and suppressing Yang can regulate the thyroid hormones, improves TCM symptoms, and decrease the adverse reaction. It can possibly regulate lymphocyte protein expression.
Adult ; Aged ; Databases, Protein ; Drugs, Chinese Herbal ; adverse effects ; pharmacology ; therapeutic use ; Electrocardiography ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Hyperthyroidism ; drug therapy ; metabolism ; pathology ; physiopathology ; Liver ; drug effects ; pathology ; physiopathology ; Lymphocytes ; drug effects ; metabolism ; Male ; Middle Aged ; Proteins ; metabolism ; Thyroid Hormones ; metabolism ; Treatment Outcome ; Yin-Yang ; Young Adult
4.Complete Genomic Sequence of Transmissible Gastroenteritis Virus TS and 3'End Sequence Characterization Following Cell Culture
Jianqiang LI ; Jie CHENG ; Xi LAN ; Xuerui LI ; Wei LI ; Xiangping YIN ; Baoyu LI ; Bin YANG ; Zhiyong LI ; Yun ZHANG ; Jixing LIU
Virologica Sinica 2010;25(3):213-224
The complete genome sequence of transmissible Gastroenteritis virus (TGEV) strain TS, previously isolated from Gansu province, was cloned and compared with published sequence data from other TGEV strains.Phylogenetic tree analysis based on the amino acid and nucleotide sequences of the S gene showed that the TGEV strains were divided into 3 clusters. TGEV TS showed a close evolutionary relationship to the American Miller cluster but had a 5' non-translated region (NTR) sequence closely related to the American Purdue cluster.Continued culture in different cell types indicated that TGEV TS virulence could be attenuated alter fifty passages in Porcine kidney (PK-15) cells, and that the Porcine kidney cell line IB-RS-2 (IBRS) was not suitable for culture of the TGEV strain TS.
5.Cloning the Structure Genes and Expression the N Gene of Porcine Epidemic Diarrhea Virus DX
Jianqiang LI ; Jixing LIU ; Xi LAN ; Jie CHENG ; Run WU ; Zhongzi LOU ; Xiangping YIN ; Xuerui LI ; Baoyu LI ; Bin YANG ; Zhiyong LI
Virologica Sinica 2009;24(3):179-186
The structure genes spike (S), nucleocapsid (N), membrane (M), small membrane (sM) of a porcine epidemic diarrhea virus (PEDV) strain DX isolated in Gansu province, North-west of China, were cloned, sequenced and compared with published sequences of PEDV strains. The nucleotide sequences encoding the entire S, sM, M and N genes open reading frame (ORF) of DX were 4 152, 231, 681 and 1 326 bases long respectively. There were transcription regulatory sequences (TRSs) upstream of the initiator ATG of the S, N and M genes. The amino acids sequences of S, M and N contained 30, 3 and 7 potential asparagine (N)-linked glycosylation sites. Homologous analysis and phylogenetic trees showed that DX had the closest relationship with strains LJB/06, JS-2004-2Z and CH/HLJH/06 that were also isolated from China and indicated the prevalence of some PEDV isolates in China were widespread since the JS-2004-2Z strain originated from the south of the China, and LJB/06 and CH/HLJH/06 were isolated from northeast China. The N gene was cloned using two primers which contained Nco I and BamH I restriction enzyme sites and subcloned into expression vector pET30a. The recombinant plasmid was then transformed into E.coli Rossta. SDS-PAGE showed there was a protein of about 55kDa as expected and Western blot indicated the N protein had biological activity.
6.Effect of depsides salts from Salvia miltiorrhiza on human hepatoma cell line SMMC-7721 subcutaneous xenografts in nude mice.
Xiangping LI ; Zhouye SONG ; Haiying ZHONG ; Zhicheng GONG ; Tao YIN ; Zanling ZHANG ; Boting ZHOU
Journal of Central South University(Medical Sciences) 2015;40(2):158-164
OBJECTIVE:
To exlpore the eff ect of depsides salts from Salvia miltiorrhiza on human hepatoma cell line SMMC-7721 xenograft tumors and the possible mechanisms.
METHODS:
A total of 36 nude mice were divided into 6 groups: A model group, a negative control group, a positive control group, and 3 treatment groups at low, middle or high dose (n=6). The tumor model of nude mice was given depsides salts at a dose of 10, 20 or 50 mg/kg every 3 day for 16 days. Then samples of subcutaneous tumors in nude mice were collected. The morphological changes of tumor samples were observed by HE staining and the expression of vascular endothelial growth factor (VEGF) and the tumor antigen Ki67 was detected by immunohistochemical method.
RESULTS:
The tumor growth was inhibited by all doses of depsides salts. The morphology of tumors was shrinkage, broken and irregularly arranged compared with the tumors in the model group and the negative control group. Morphological changes were more obvious in tumors with treatment at high dose. Expression of VEGF and Ki67 in treatment groups and the positive control group were lower than that in the model group and the negative control group, with a significant difference (P<0.05).
CONCLUSION
Depsides salts from Salvia miltiorrhiza can inhibit the growth of human hepatoma cell line SMMC-7721 tumor in nude mice, which is related to the inhibition of Ki67 and VEGF.
Animals
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Carcinoma, Hepatocellular
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pathology
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Cell Line, Tumor
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drug effects
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Depsides
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pharmacology
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Humans
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Ki-67 Antigen
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metabolism
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Liver Neoplasms
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pathology
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Mice
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Mice, Inbred BALB C
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Mice, Nude
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Salts
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Salvia miltiorrhiza
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chemistry
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Vascular Endothelial Growth Factor A
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metabolism
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Xenograft Model Antitumor Assays
7.The design and clinical application of a multidisciplinary round electronic checklist in ICU
Xiangping CHEN ; Yin LIANG ; Yiyu ZHUANG ; Feng GUO ; Yueliang CHEN ; Yinshan WU ; Limin HUANG ; Luoyang TU
Chinese Journal of Practical Nursing 2018;34(9):672-676
Objective To investigate the effects of a multidisciplinary round electronic checklist in ICU. Methods From October 2016 to September 2017,a total of 1 172 patients were provided with a multidisciplinary round electronic checklist. Another 886 patients with routine rounds between October 2015 to September 2016 were retrospectively analyzed. The duration of mechanical ventilation and ICU stay,the rate of nosocomial infection, the percentages of catheter days(central line catheter days,artificial airway days and urinary catheter days) and clinical staffs' satisfaction with the multidisciplinary collaboration were compared. Results The duration of mechanical ventilation and ICU stay in the intervention group were 1.42(0.68,3.63)days and 3.19(2.48,5.40)days respectively,in the control group were 2.16(1.40, 4.77) days and 4.14(3.46, 6.92) days respectively. The differences between two groups were statistically significant(t=2.084, 2.247, P <0.05). The application of a multidisciplinary round electronic checklist shortened the duration of mechanical ventilation and ICU stay. The percentages of catheter days and the rate of catheter-associated urinary tract infection were 0(0/6 269),60.34%(6 269/10 389)respectively,in the control group were 0.93‰(6/6 485),67.02%(6 485/9 676)respectively.The differences between two groups were statistically significant(χ2=96.187,Fisher's exact test=0.031,P<0.05).Besides,the level of multidisciplinary collaboration in ICU was greatly improved in the intervention group, 3.03 ± 0.11 vs. 4.13 ± 0.39 (t=24.587, P < 0.05). Conclusions A multidisciplinary round electronic checklist in ICU can improve the care quality and multidisciplinary collaboration.
8.Survey on the knowledge, attitude, and practices of breastfeeding among doctors and nurses in the neonatal intensive care unit of Qianxinan Prefecture, Guizhou Province
Chunjiang CHEN ; Shunfen WU ; Lu ZENG ; Liqing WU ; Xiangping KONG ; Hao YIN ; Yi ZHANG ; Zhu ZHU ; Shixia WANG ; Wanbin GOU ; Guangjie WEI
Chinese Journal of Perinatal Medicine 2024;27(7):553-561
Objective:To understand the breastfeeding situation in the neonatal intensive care units (NICUs) in Qianxinan Prefecture, Guizhou Province, and to assess the knowledge, attitudes, and practices of doctors and nurses regarding breastfeeding, aiming to provide foundational data for improving breastfeeding quality.Methods:A questionnaire was developed to survey the knowledge, attitudes, and practices related to breastfeeding in NICUs. The questionnaire was divided into three dimensions: knowledge (seven items, total score of 7), attitudes (nine items, total score of 45), and practices (seven items, total score of 35). Lower scores indicated weaker recognition of breastfeeding. Additionally, five items were included to identify the most influential factors affecting breastfeeding. From November 25 to November 30, 2023, a survey was conducted among doctors and nurses with professional qualifications who had worked in the neonatal departments of nine hospitals in Qianxinan Prefecture for at least one year. Independent sample t-tests and Chi-square tests were used to compare the scores of doctors and nurses from different levels of hospitals and within the same level of hospitals across the three dimensions. Results:(1) Among the nine hospitals, three were tertiary grade A hospitals (referred to as "tertiary hospitals"), with 95.6% (43/45) of the doctors and 96.5% (110/114) of the nurses participating in the survey. Six were secondary grade A hospitals (referred to as "secondary hospitals"), with 95.0% (38/40) of the doctors and 97.6% (83/85) of the nurses participating. (2) All nine hospitals were baby-friendly hospitals and all had breastfeeding promotional materials. Six hospitals had NICUs that promoted breastfeeding, with an average NICU breastfeeding rate of 25.8% across the prefecture between year 2021 to 2023. (3) The proportion of doctors who had received breastfeeding training was higher than that of nurses within the same level of hospitals [tertiary hospitals: 69.8% (30/43) vs. 40.0% (44/110), χ 2=10.97, P=0.001; secondary hospitals: 47.4% (18/38) vs. 24.1% (20/83), χ 2=6.55, P=0.010], although the overall training rates were low. (4) In tertiary hospitals, doctors scored higher than nurses in the attitude dimension [(35.35±4.75) vs. (33.18±5.60) scores, t=-2.03, P=0.044] and also in the practice dimension [(26.98±3.00) vs. (25.60±3.75) scores, t=-2.17, P=0.032]. In secondary hospitals, the total knowledge dimension score of doctors was higher than that of nurses [(4.92±1.44) vs. (4.20±1.45) scores, t=-2.52, P=0.013]. In tertiary hospitals, the total scores for attitude and practice dimensions of doctors were higher than those of doctors in secondary hospitals, and the total scores for knowledge, attitude, and practice dimensions of nurses were higher than those of nurses in secondary hospitals (all P<0.05). (5) In the knowledge dimension, the lowest scoring item of doctors in the tertiary hospitals was "Breastfeeding is possible for maternal hepatitis B newborns after receiving vaccines and immunoglobulin"; the lowest scoring item of nurses in the tertiary hospital, and doctors and nurses in the secondary hospitals was "The duration of breastfeeding has a greater impact on neonatal outcomes". In the attitude dimension, the lowest scoring item for doctors and nurses in both tertiary and secondary hospitals was "You think the breastfeeding process is more troublesome than feeding preterm formula". In the practice dimension, the lowest scoring item of the doctors and nurses in the tertiary hospitals was "Your hospital had enough breastfeeding knowledge training", while for the doctors and nurses in the secondary hospitals were "You have more opportunities to participate in various breastfeeding-related training" and "Breast feeding should be started as soon as possible when the infant is stable after active treatment", respectively. (6) The most influential factors affecting breastfeeding were: lack of cooperation from parents (50.0%, 137/274), relative insufficient human resources for doctors and nurses (21.9%, 60/274), and the absence or poor implementation of breastfeeding management policies (18.3%, 50/274), etc. Conclusions:The breastfeeding rate in NICU of county-level hospitals is relatively low, and medical staff, especially nurses, have insufficient knowledge about breastfeeding. It is necessary to strengthen various breastfeeding training for medical staff to enhance their understanding of NICU breastfeeding.
9.Establishment and application of evaluation criteria for clinical rational drug use of atosiban based on AHP-TOPSIS method
Jinjin YIN ; Peili DU ; Xiangping TAN ; Zhengrong MEI ; Shengying SHI ; Shaozhi LIU
China Pharmacy 2023;34(1):86-90
OBJECTIVE To establish evaluation criteria for rational drug use of atosiban in clinic, and to provide reference for rational drug use of atosiban in clinic. METHODS Based on the drug instructions of atosiban acetate injection and related guidelines, the experts of the Evaluation Group of Rational Drug Use formulated the evaluation criteria of rational drug use, including 5 primary indexes and 8 secondary indexes. The weight coefficients of secondary indexes were calculated by analytic hierarchy process (AHP), and the use of atosiban acetate injection in 190 pregnant women from the Third Affiliated Hospital of Guangzhou Medical University (referred to as “our hospital”) was evaluated retrospectively by technique for order preference by similarity to an ideal solution (TOPSIS). The evaluation results were divided into three levels including reasonable, basic reasonable and unreasonable application based on the relative approach degree. RESULTS Among 190 pregnant women, 49 (25.8%) were treated with atosiban reasonably, 39 (20.5%) were treated with atosiban basic reasonably, and 102 (53.7%) were treated with atosiban unreasonably. The evaluation results obtained by AHP-TOPSIS method were consistent with the actual situation in clinic. The main problems of the unreasonably use were super indications, unreasonable usage and dosage, over the course of treatment and the lack of proper economic consideration. CONCLUSIONS The rationality evaluation criteria of atosiban’s clinical application are established by AHP-TOPSIS method; the evaluation results obtained by this method are quantifiable, scientific and reliable. The unreasonable use of atosiban is common in our hospital, and the management should be strengthened in clinical application.
10.Genome-wide analysis identify novel germline genetic variations in ADCY1 influencing platinum-based chemotherapy response in non-small cell lung cancer.
Chenxue MAO ; Juan CHEN ; Ting ZOU ; Yuankang ZHOU ; Junyan LIU ; Xi LI ; Xiangping LI ; Min LI ; Pinhua PAN ; Wei ZHUO ; Yang GAO ; Shuo HU ; Desheng XIAO ; Lin WU ; Zhan WANG ; Heng XU ; Wen YANG ; Yingjie XU ; Haihua XIAO ; Kazuhiko HANADA ; Wei ZHANG ; Honghao ZHOU ; Jiye YIN ; Zhaoqian LIU
Acta Pharmaceutica Sinica B 2022;12(3):1514-1522
To explore the pharmacogenomic markers that affect the platinum-based chemotherapy response in non-small-cell lung carcinoma (NSCLC), we performed a two-cohort of genome-wide association studies (GWAS), including 34 for WES-based and 433 for microarray-based analyses, as well as two independent validation cohorts. After integrating the results of two studies, the genetic variations related to the platinum-based chemotherapy response were further determined by fine-mapping in 838 samples, and their potential functional impact were investigated by eQTL analysis and in vitro cell experiments. We found that a total of 68 variations were significant at P < 1 × 10-3 in cohort 1 discovery stage, of which 3 SNPs were verified in 262 independent samples. A total of 541 SNPs were significant at P < 1 × 10-4 in cohort 2 discovery stage, of which 8 SNPs were verified in 347 independent samples. Comparing the validated SNPs in two GWAS, ADCY1 gene was verified in both independent studies. The results of fine-mapping showed that the G allele carriers of ADCY1 rs2280496 and C allele carriers of rs189178649 were more likely to be resistant to platinum-based chemotherapy. In conclusion, our study found that rs2280496 and rs189178649 in ADCY1 gene were associated the sensitivity of platinum-based chemotherapy in NSCLC patients.