2.Cloning of Acidic Xylanase Gene and Its Secretion Expression in Pichia pastoris
Chun-Hua LI ; Xiang LI ; Li-Xin MA ;
Microbiology 1992;0(06):-
An acidic xylanase gene,named xyl3,was cloned from the genomic library of enviromental microbes constructed by using shotgun cloning strategy,and submitted to GeneBank with accession number of gb:AY300805.BLAST analysis indicated that the gene xyl3 has low similarity with other xylanase genes and the encoded xylanase,sorted as Glycosyl hydrolases family 10,has 77% similarity with the intra-cellular xylanase from Geobacillus stearothermophilus at amino acid level.Treated with T4 DNA polymerase,the gene xyl3 was ligated with the linearized Pichia pastoris expression vector pHBM905 produced by digestion of restriction endonuclease CpoI and NotI to generate the recombinant plasmid pHBM706.Then the plasmid pHBM706,digested by restriction endonuclease SalI,was transformed into P.pastoris GS115 to obtain the recombinant P.pastoris GS115(pHBM706),which was induced to produce the recombinant xylanase with 0.5% methanol at 28℃.At the 36th hours of induction,the porduced crude enzyme was detected to reach the higest enzyme activity of 0.177 IU/mL.The optimal pH and temperature of the enzyme activity is 5.5 and 50℃ respectively.
3.Survey on Sleep Time in Korean Children
xiu-fang, HAO ; yan-xiang, MA ; hong-hua, LIU
Journal of Applied Clinical Pediatrics 2006;0(22):-
Objective To evaluate the sleep characteristics of the Korean children,and explore the measurement of improving the sleep qua-lity of minority nationality children.Methods The sleep time of 1 183 from 3 to 12 years age Korean children were investigated with questionnaire The sleep time of korean children in different ages and cities were compared.Results The mean time of total sleep time in Korean children was(10.06?1.29) h,which was decreasing with the age′s increasing.The difference in different ages groups was remarkable(Pa
4.Pharmacokinetics of loganin, ferulic acid and stilbene glucoside in Bushen Tongluo formula in vivo.
Xiang-dan LIU ; Pan HUANG ; Yue-hua LU ; Ming MA ; Ri-bao ZHOU ; Lin-xiang YUAN ; Xin-jun PENG
China Journal of Chinese Materia Medica 2015;40(12):2428-2434
To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral
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Animals
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Biological Availability
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Coumaric Acids
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administration & dosage
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blood
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pharmacokinetics
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Drugs, Chinese Herbal
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administration & dosage
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analysis
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pharmacokinetics
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Glucosides
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administration & dosage
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blood
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pharmacokinetics
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Iridoids
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administration & dosage
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blood
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pharmacokinetics
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Male
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Rats
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Rats, Sprague-Dawley
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Stilbenes
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administration & dosage
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blood
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pharmacokinetics
5.Unilateral fixation combined with interbody fusion for the treatment of lumbar degenerative instability via a paraspinal intermuscular approach under MAST Quadrant retractor.
Sheng-hua HE ; Du-jun MA ; Xiang ZHAO ; Hao DING ; Jun FANG
China Journal of Orthopaedics and Traumatology 2016;29(5):424-428
OBJECTIVETo study clinical effects of unilateral pedicle screw-rod system fixation combined with the single the cage of interbody fusion in the treatment of lumbar degenerative instability via paraspinal intermuscular approach under MAST Quadrant retractor.
METHODSFrom February 2010 to December 2011, 39 patients with lumbar degenerative instability after invalid conservative treatment were treated with unilateral fixation combined with interbody fusion via the paraspinal intermuscular approach under MAST Quadrant retractor. The indexes such as the operative time, blood loss, complications, VAS pain scores, JOA (subjective symptoms of low back pain,lower limb pain and numbness, sensory disturbance, movement disorders) scores, modified Macnab criteria for curative effect evaluation were observed before treatment, at the 1st month after treatment and the latest follow-up. The intervertebral fusion was studied on the X-ray at the latest follow-up.
RESULTSAll the patients were followed up, and the mean during was (22.3 +/- 8.6) months. The operation time was (138 +/- 46) min, and the amount of bleeding was (335 +/- 152) ml. There were no complications such as cerebrospinal fluid leakage and spinal nerve injury during operation, and no incision infection after operation. The VAS pain score was reduced from preoperative 7.93 +/- 1.27 to 2.05 +/- 1.18 on the 1st month after operation and 1.89 +/- 0.42 at the latest follow-up. The JOA total score was improved from preoperative 1.59 +/- 0.42 to 8.86 +/- 0.37 on the 1st month and 9.02 +/- 0.29 at the latest follow-up. According to modified Macnab criteria, there were 17 cases got an excellent result, 19 good, 3 case fair. Thirty-five patients got intervertebral bony fusion at the latest follow-up.
CONCLUSIONUnilateral pedicle screw-rod system fixation combined with single cage interbody fusion in the treatment of lumbar degenerative instability via paraspinal intermuscular approach under MAST Quadrant retractor is a safe, minimally invasive, satisfactorily effective methods to treat lumbar degenerative instability.
Adult ; Aged ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Intervertebral Disc Degeneration ; surgery ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Reconstructive Surgical Procedures ; Spinal Fusion ; Treatment Outcome
6.Study of atria-His bundle sequential pacing on cardiac electrophysiology and heamodynamics in dogs.
Chinese Journal of Applied Physiology 2002;18(1):71-74
AIMTo evaluate the effects of atria-His bundle sequential pacing on cardiac electrophysiology and heamodynamics in dogs.
METHODSIn 20 opening chest anesthetized dogs, platinum electrodes were fixed at the epicardium of right atria (RA) and the right ventricular apex (RVA) respectively, pacing right atria and the right ventricle. A special lead was located at His bundle (based on a optical "H" wave and narrow duration of the QRS complexes recorded in ECG), pacing His bundle. Cardiac electrophysiology and hemodynamics parameters were compared in the different pacing models RA(AAI, RVA-(VVI), HisB-(VVI) single chamber pacing and RA-RVA(DDI), RA-HisB(DDI) dual chamber pacing.
RESULTSThe threshold of His B pacing is similar to that of RVA pacing. Cardiac output (CO) is increased in pacing of RA(AAI), His B-(VVI) and RA-His B(DDI). It is increased by 29.64% in pacing of RA-His B(DDI) (P < 0.01) and by 0.25% (P > 0.05) in pacing of RA-RVA(DD1) While CO is decreased by 5.41% in RVA-VVI) pacing (P > 0.05). SV, LVSW and RVSW of RA-HisB(DDI) pacing are superior to those in RVA-VVI) and RA-RVA(DDI) pacing.
CONCLUSIONRight atria-His bundle sequence pacing significantly improves cardiac function compared with the other model pacing because it maintains normal physiological electronic activity sequence and systolic synchrony. It will be adapted to clinical application.
Animals ; Bundle of His ; physiology ; Cardiac Electrophysiology ; Cardiac Pacing, Artificial ; methods ; Dogs ; Female ; Heart Atria ; Hemodynamics ; Male
7.Expression of Hox transcript antisense RNA, enhancer of zeste homolog 2 and vascular endothelial growth factor in nasopharyngeal carcinoma tissues and their correlation with prognosis
Yun MA ; Hua ZOU ; Xiang LIU ; Peng TIAN ; Yan NIE ; Shaohua QU
Chinese Archives of Otolaryngology-Head and Neck Surgery 2017;24(5):228-232
OBJECTIVE To explore the expression of long non-coding RNA (LncRNA) HOTAIR, enhancer of Zeste homolog 2 (EZH2) and vascular endothelial growth factor (VEGF) in nasopharyngeal carcinoma (NPC) and their relationship between HOTAIR and prognosis. METHODS We examined the expression of HOTAIR, EZH2 and VEGF in NPC tissues, and analyzed the clinical significance of HOTAIR expression in patients with NPC. RESULTS We found that the expression of HOTAIR, EZH2 and VEGF in the NPC tissues were significantly higher than those in the chronic nasopharyngitis tissues in the gene and protein levels (U=955,P<0.05). The HOTAIR was positively correlated with VEGF (r=0.599,P<0.001), VEGF was positively correlated with EZH2 (r=0.369, P=0.012), and HOTAIR had no significant correlation with EZH2 (r=0.139,P=0.357). Moreover, HOTAIR expression levels increased with clinical stage progression. CONCLUSION The expression of HOTAIR, EZH2 and VEGF were significantly increased in the NPC tissues, and the expression of HOTAIR was related with the TNM stage, the clinical stage and the short term therapeutic effect of the NPC patients, which indicated that the HOTAIR might be used as a biological indicator to determine the prognosis of the NPC.
8.Effects of Zibu Piyin Recipe on Protein Expression of PDHE1α in Peripheral Tissues and Brain of Rats with Spleen Yin Deficiency Diabetes
Lina LIANG ; Wei MA ; Libin ZHAN ; Shouyu HU ; Luping ZHENG ; Hua SUI ; Hong XIANG
Chinese Journal of Information on Traditional Chinese Medicine 2014;(11):52-55
Objective To explore the mechanism of Zibu Piyin Recipe (ZBPYR) on spleen yin deficiency diabetes-associated cognitive disorder (DACD). Methods The rats were randomly divided into control group, diabetes mellitus (DM) group, spleen yin deficiency group, spleen yin deficiency DM group and spleen yin deficiency DM+ZBPYR group (treatment group). Type 2 DM models were established by high-fat food feeding and low dose STZ intraperitoneal injection for 4 weeks. Then the classical compound method was used to construct spleen yin deficiency rat models by improper diet, over exertion and yin fluids exhaustion. The treatment group was given ZBPYR by gavage for 15 days, and the other groups were given the same amount of normal saline. Then cerebral cortex, hippocampus, stomach and liver were obtained and the changes of protein expression of PDHE1α in them were observed by Western Blot. Results The protein expression of PDHE1αin cortex of DM group and spleen yin deficiency DM group were lower than control group (P<0.05). PDHE1α expression of treatment group in cortex and stomach increased more significantly than spleen yin deficiency DM group (P<0.05). The expression of PDHE1α protein showed no significant difference among all groups in hippocampus and liver. Conclusion ZBPYR improved spleen yin deficiency DACD by regulating PDHE1αin cortex and stomach.
9.Effects of tonifying spleen and promoting blood circulation on the retina and Bruch membrane in apolipoprotein E-deficient mouse with dyslipidemia
Yi, WANG ; Yu-lan, MA ; Juan, LI ; Qing-hua, ZENG ; Luo-xiang, LI
Chinese Journal of Experimental Ophthalmology 2013;(1):13-17
Background Dyslipidemia is one of the major causes of age-related macular degeneration (AMD).At present,the study of the preventive and treating methods of A MD is still a hot spot.Objective The purpose of this study was to observe the effect of tonifying the spleen and promoting blood circulation on the retina and Bruch membrane in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Thirty-six ApoE-/-mice aged 2 months were randomly divided into the normal diet group,high fat diet group and medicine group.A diet with a higher content of fat was given for 5 consecutive months to the mice of the high fat diet group and medicine group,and in the last month,a concoction that tonifies the spleen and promotes blood circulation was gavagely administered in the medicine group,and an equivalent volumes of normal saline solution was administered in the same way in the normal diet group and high fat diet group.Total plasma cholesterol (TC),low density lipoprotein (LDL)and triglyceride (TG) were detected by (ELISA? Name of assay?) using the 7170 Hitachi automatic biochemical analyzer,and morphological changes of the retina and Bruch membrane were examined by light microscopy and transmission electron microscopy.The number of outer nuclear layer (ONL) cells,retinal pigment epithelial (RPE) cells and the thickness of the Bruch membrane were examined by semi-quantitative histopathology with the Mias 2000 Imaging Analyzer System.Results The concentrations of TC,LDL and TG were (6.47 ±0.49) mmol/L,(1.46 ±0.10)mmol/L and (0.62 ±0.21) mmol/L,respectively,in 7-month-old mice of the medicine group,showing a significant reduction in comparison with (10.53 ±0.30) mmol/L,(1.90±0.13) mmol/L,(1.15±0.29) mmol/L of the high fat diet group,and (9.63 ± 0.18) mmol/L,(1.12 ± 0.15) mmol/L,(0.88 ± 0.21) mmol/L in the normal diet group (P<0.05-0.01).The disorder and atrophy of ONL and RPE cells,divergence of fiber of the Bruch membranes were found in both the high fat diet group and normal diet control group under the light microscope,and drusen formed in some of the mice in the high fat diet group.However,ONL and RPE were well organized in the medicine group.The cell numbers in the ONL and RPE layer in the 7-month-old mice were (23 124.00±755.18) and (10.75±0.59),respectively,in the medicine group,(19 107.00 ± 1436.82) and (8.55 ± 1.11),respectively,in the high fat diet group,(21 663.00± 1073.27) and (9.75 ±0.58),respectively,in the normal diet group,with significant differences among them (P<0.05-0.001).Thickness of the Bruch membrane in the medicine group extensively reduced in high fat diet group and normal diet control group (P<0.01).The ultrastructures of the RPE and Bruch membrane were much more improved in the mdedicine group.Conclusions Tonifying the spleen and promoting blood circulation can attenuate hyperlipemia in ApoE-/-mouse;furthermore,it lessens the pathological abnormalities in the ONL,RPE and Bruch membrane.
10.Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites
Hong-Bing MA ; Yu-Hua QIU ; Ran TAO ; Wen-Xiang LI ; Ying XU ; Xue-Guang ZHANG ;
China Biotechnology 2006;0(08):-
A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.