1.3 years follow-up of refractory systemic lupus erythematosus patients with transplantation treating of umbilical cord mesenchymal stem cells
Ru BAI ; Yan QI ; Zhaoping Lü ; Mingyao MENG ; Jinwen WANG ; Zongliu HOU
Chinese Journal of Immunology 2017;33(6):905-909
Objective:To observe immunological indexes,the quantity of cytokine expression and clinical curative effect of umbilical cord mesenchymal stem cells between before and after the treatment of systemic lupus erythematosus patients.Methods:Selected 10 cases of SLE,on the basis of glucocorticoid and immune inhibitor treatment,intravenous injection UC-MSC of cultivating proliferation within 6 generations.Before and after treatment of UC-MSC testing the relative quantity of cytokine of CTLA-4,IL-15,IL-2,CD86,IL-17c,Foxp3,TGF-β2 which were related of immunopathogenesis of SLE.Before and after treatment to determined SLE disease activity index (SLEDAI) score and detection of blood in the urine routine,liver and kidney function,24 hours urinary protein quantitative,immunoglobulin and complement levels.Results:After treatment the relative expression value of IL-15 and IL-2 was decreased,CTLA-4 was risen.There had no significant difference with the relative expression value of CD86,IL-17c,Foxp3,TGF-β2 in before and after treatment of UC-MSC.After treatment serum complement C3 and C4 level,serum albumin,were risen.24 hour proteinuria and SLEDAI were decreased.There was no serious adverse reaction occurred,no complications related to transplantation in 10 cases.Conclusion:UC-MSC can regulate the expression of cytokines of participate in the immune response in the patients with SLE.Treatment of SLE by UC-MSC can elevate serum albumin and C3 and C4 level,reduce the 24 hours urinary protein quantity,relife kidney damage,improve clinical symptoms;UC-MSC transplantation in patients with SLE have good security;UC-MSC transplantation may be a feasible method for the treatment of SLE.
2.Synthesis and anti-tumor activity of ursolic acid derivatives.
Yan-qiu MENG ; Dan LIU ; Zhong-wei BAI ; Ling-li CAI ; Hong-ru AI
Acta Pharmaceutica Sinica 2011;46(5):556-560
Structure of natural product-ursolic acid was modified for increasing its antitumor activity. Ursolic acid was acylated, esterified, hydrolized or oxidized to obtain target pentacyclic triterpenoid compounds with different substitutes. Sixteen derivatives of ursolic acid were designed and synthesized including eleven new compounds. Anti-tumor activities of ursolic acid and these derivatives against HeLa, SKOV3 and BGC-823 cells in vitro were investigated by MTT assay. The results indicated that compounds 7a and 8a were found to have stronger cell growth inhibitory than ursolic acid on HeLa cells and SKOV3 cells separately, and are worth to be intensively studied further.
Antineoplastic Agents, Phytogenic
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chemical synthesis
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chemistry
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pharmacology
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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HeLa Cells
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Humans
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Triterpenes
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chemical synthesis
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chemistry
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pharmacology
3.Improvement of the thermostability of xylanase by N-terminus replacement.
Hao-Meng YANG ; Kun MENG ; Hui-Ying LUO ; Ya-Ru WANG ; Tie-Zheng YUAN ; Ying-Guo BAI ; Bin YAO ; Yun-Liu FAN
Chinese Journal of Biotechnology 2006;22(1):26-32
The hybrid xylanase TB was constructed by the substitution of the N-terminus segment of the Streptomyces olivaceoviridis xylanase XYNB with corresponding region of Thermomonosporafusca xylanase TfxA. The hybrid gene tb, encoding the TB, was correctly expressed in Escherichia coli BL21 and Pichia pastoris GS115. TB was purified and its enzymatic properties were determined. The results revealed that the optimal temperature and optimal pH of TB were at 70 degrees C and 6.0, which have been obviously improved compared with those of XYNB. The thermostability of TB were all about six-fold of XYNB's after incubating the properly diluted enzyme solutions at 80 degrees C and 90 degrees C for 3min, respectively. The pH stability of TB was 5 to approximately 9, which was narrower than that of XYNB. Still, TB remains a high specific activity as XYNB does. Analysis of a homology modeling and sequence similarity were used to reveal the factors influencing the enzymatic properties of TB and the discussion for the relationship between structure and function of xylanase was given.
Amino Acid Sequence
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Base Sequence
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Desulfurococcaceae
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enzymology
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genetics
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Endo-1,4-beta Xylanases
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genetics
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metabolism
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Enzyme Stability
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Escherichia coli
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enzymology
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genetics
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Hot Temperature
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Molecular Sequence Data
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Pichia
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enzymology
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genetics
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Protein Engineering
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methods
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Recombinant Fusion Proteins
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genetics
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metabolism
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Streptomyces
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enzymology
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genetics
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Structure-Activity Relationship
4.Umbilical hernia repair in conjunction with abdominoplasty.
Ming BAI ; Meng-Hua DAI ; Jiu-Zuo HUANG ; Zheng QI ; Chen LIN ; Wen-Yun DING ; Ru ZHAO
Chinese Journal of Plastic Surgery 2012;28(5):349-351
OBJECTIVETo investigate the feasibility and clinical benefits of umbilical hernia repair in conjunction with abdominoplasty.
METHODSThe incision was designed in accord with abdominoplasty. The skin and subcutaneous tissue was dissected toward the costal arch, and then the anterior sheath of rectus abdominus was exposed. After exposure and dissection of the sac of umbilical hernia, tension-free hernioplasty was performed with polypropylene mesh. After dissecting the redundant skin and subcutaneous tissue, the abdominal wall was tightened.
RESULTSBetween May 2008 and May 2011, ten patients were treated in the way mentioned above. The repair of umbilical hernia and the correction of abdominal wall laxity were satisfactory. There was no recurrence of umbilical hernia, hematoma, seroma or fat liquefaction.
CONCLUSIONThrough careful selection of patients, repair of umbilical hernia and body contouring could be achieved simultaneously.
Abdominal Wall ; surgery ; Abdominoplasty ; methods ; Adult ; Female ; Hernia, Umbilical ; surgery ; Humans
5.Improving phytase expression by increasing the gene copy number of appA-m in Pichia pastoris.
Hui-Ying LUO ; Huo-Qing HUANG ; Ying-Guo BAI ; Ya-Ru WANG ; Pei-Long YANG ; Kun MENG ; Tie-Zheng YUAN ; Bin YAO
Chinese Journal of Biotechnology 2006;22(4):528-533
In order to improve the fermentation potency of phytase in recombinant host and decrease the production cost, the pichia expression vector pGAPZalpha-A was modified by introduction of an AOX1 promoter from vector pPIC9 and the resulted vector pAOXZalpha is an methanol induced vector. After that, a phytase gene appA-m was cloned into pAOXZalpha to construct the recombinant vector pAOXZalpha-appA-m. The recombinant Pichia pastoris 74#, which already contains one copy of appA-m and its fermentation potency exceeded 7.5 x 10(6) IU/mL, was used as the host strain for the transformation of pAOXZalpha-appA-m. The Pichia pastoris transformants were gained by electroporation. PCR results indicated that the appA-m expression box has integrated into the genome of Pichia pastoris and the original construction of phytase gene has not changed. SDS-PAGE analysis revealed that phytase was overexpressed and secreted into the medium supernatant. Recombinants with high expression level were screened and used for fermentation. In 5L fermentor, the expression level of phytase protein achieved 4 mg/mL and the phytase activity (fermentation potency) exceeded 1.2 x 10(7) IU/mL, which was about 1.6-fold compared with that of the host strain 74#. Moreover, the improved recombinant Pichia pastoris is excellent at expression stability and heredity stability.
6-Phytase
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genetics
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Fermentation
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Gene Dosage
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Pichia
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genetics
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Plasmids
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Polymerase Chain Reaction
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Recombination, Genetic
6.Hydrophobic interaction between beta-sheet B1 and B2 in xylanase XYNB influencing the enzyme thermostability.
Hao-Meng YANG ; Bin YAO ; Hui-Ying LUO ; Wang-Zhao ZHANG ; Ya-Ru WANG ; Tie-Zheng YUAN ; Ying-Guo BAI ; Ning-Feng WU ; Yun-Liu FAN
Chinese Journal of Biotechnology 2005;21(3):414-419
A homology modeling of xylanase XYNB from Streptomyces olivaceoviridis A1 was made by Swiss-Model. The hydrophobic Interaction between beta-sheet B1 and B2 in the tertiary structure model of XYNB was compared with other thermophilic xylanase. A T11Y mutation was introduced in XYNB by site-dirrected mutagenesis to improve the thermostability of the enzyme. The XYNB and mutant xylanase (XYNB') expressed in Pichia pastoris were purified and their enzymatic properties were determined. The result revealed that the thermostability of XYNB' was obviously higher than that of XYNB. The optimal temperature of XYNB' for its activity was 60 degrees C, similar to XYNB. But, compare to XYNB, the optimal pH value, the Km value and the specific activity of XYNB' had also been changed. The research results suggested that the aromatic interaction between beta-sheet B1 and B2 in xylanase should increase enzyme thermostability. The mutant xylanase XYNB' is a good material for further research in the relationship between structure and function of xylanase.
Bacterial Proteins
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chemistry
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genetics
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Endo-1,4-beta Xylanases
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chemistry
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genetics
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Enzyme Stability
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Hot Temperature
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Hydrophobic and Hydrophilic Interactions
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Mutagenesis, Site-Directed
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Mutant Proteins
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chemistry
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Pichia
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genetics
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metabolism
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Protein Conformation
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Protein Folding
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Protein Structure, Tertiary
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genetics
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Recombinant Proteins
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biosynthesis
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genetics
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Streptomyces
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enzymology
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genetics
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beta-Glucosidase
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chemistry
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genetics
7.Development and application of a new digital electric fire needle instrument.
Meng-Ru HE ; Xin HUI ; Bai-Xiao ZHAO ; Ze-Lin CHEN
Chinese Acupuncture & Moxibustion 2023;43(5):593-596
In view of the limitations of the high operational difficulty, safety hazards and adverse reactions of traditional fire needle, and unclear treatment parameters of existing electric fire needles, a new digital electric fire needle instrument was designed and developed in this study. This instrument is a gun type structure, consisting of a gun body, a power supply interface on the gun body, a display unit and a drive unit, a heating unit, a cooling unit, a positioning unit, and a needle inserting unit in the gun body. This instrument can digitally realize the regulation of parameters such as fire needle inserting temperature, depth and speed, and it has the advantageous features of intelligent needle burning, precise positioning, and safe and easy operation. This instrument meets the needs of more patients, medical professionals and scientific researchers, and is conducive to promoting the development of fire needle acupuncture therapy.
Humans
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Needles
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Heating
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Research Personnel
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Temperature
8.Development of a GeXP based multiplex RT-PCR assay for simultaneous differentiation of nine human hand food mouth disease pathogens.
Xiu-Mei HU ; Yong ZHANG ; Bang-Lao XU ; Meng-Jie YANG ; Miao WANG ; Chen ZHANG ; Jin LI ; Ru-Yin BAI ; Xiao-Mian ZHOU ; Wen-Bo XU ; Xue-Jun MA
Chinese Journal of Virology 2011;27(4):331-336
A multiplex RT-PCR assay based on GeXP system was developed in order to detect simultaneously human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) and other coxsackieviruses (CVA4, 5, 9 and 10, CVB1, 3 and 5). Enterovirus detection was performed with a mixture of 12 pairs of oligonucleotide primers including one pair of published primers for amplifying all known pan-enterovirus genomes and eleven primer pairs specific for detection of the VP1 genes of EV71, C A16, CVA4, CVA5, CVA9, CVA10, CVB1, CVB3 and CVB5, respectively. The specificity of multiplex RT-PCR system was examined using enterovirus cell cultures and positive strains identified previously from hand-foot-and-mouth disease (HFMD) patients. Serial dilution of titrated EV71 and C A16 cell cultures and in vitro transcripted RNA of enterovirus VP1 regions were used to detect the sensitivity of the multiplex RT-PCR system. The limit of detection for this multiplex RT-PCR system was 10(0.5) TCID50/microL for EV71 and C A16 cell cultures and 1000 copies for in vitro transcripted RNA of nine viruses per assay. This multiplex RT-PCR assay is a rapid, sensitive and specific assay for the diagnosis of common enterovirus infection in cases of HFMD outbreak and is also potentially useful for molecular epidemiological investigation.
DNA Primers
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genetics
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Enterovirus
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classification
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genetics
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isolation & purification
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Hand, Foot and Mouth Disease
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diagnosis
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virology
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Humans
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Reverse Transcriptase Polymerase Chain Reaction
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methods
9.Expression and regulation of drug transporters in placenta
Meng-ru BAI ; Dong-li SUN ; Hui-di JIANG ; Cai-hong ZHENG
Acta Pharmaceutica Sinica 2016;51(6):879-
Placenta, an important organ, mediates the exchange of nutrients and metabolites between mother and fetus. The transporters, including ATP-binding cassette (ABC) transporters and solute carrier (SLC), expressed in the syncytiotrophoblast play a vital role in substance exchange. Some transporters, such as organic cation transporters (OCTs) and organic anion transporters (OATs), mediate the uptake of endogenous substances and drugs. Some transporters, such as P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and multidrug resistance-associated proteins (MRPs), can excrete their substrates from the syncytiotrophoblast to the maternal circulation. However, the expression and activity of these transporters are not uniform throughout the gestation period, since they can be affected by physiological and pathological changes during pregnancy or drugs. Thus, an understanding of the role of placental transporters and the variation in their expression and activity in response to physiological and pathological changes is essential for efficient and safe therapy during pregnancy, and it also has important value in the development of drug treatment in pregnancy.