6.Cloning Chitinase Gene of the Entomopathogene Fungus Metarhizium anisopliae HN1 and High-level Expression in Escherichia coli
Wen-Bin REN ; Shi-Qing ZHANG ; Jun-Sheng HUANG ;
China Biotechnology 2006;0(07):-
Chitinases genes from Metarhizium anisopliae which is an important entomopathogenic fungus were considered one of the key factors to invade their hosts. One Metarhizium anisopliae HN1 strain was isolated and screened. A chitinase gene was amplified by RT-PCR from Metarhizium anisopliae HN1, The whole length of this gene was 1275bp,and the nucleotide sequence of the gene was 96% similarity to that of the M. anisopliae E6 accessed in GenBank ( AF02749). The gene has been registered in GenBank and its accession number is DQ011865. The gene was subcloned into prokaryon expression vector pET-22b( + ), transformed this recombinant expression plasmid into E. coli strain BL 21 and effective expressed. The SDS-PAGE analysis indicated that the recombinant protein was 42kDa which is same to the reported article. The expression level of recombinant protein was about 63. 3% of whole expressed proteins , And when recombinant E. coli were crushed by freeze and supersonic wave , the activity assay indicates that the chitinase expressed in bacteria possesses biological activity.
7.Biomechanical effects of osteoporosis vertebral augmentation on the adjacent intervertebral levels using a three-dimensional finite element analysis
Sheng LU ; Yong-Qing XU ; Mei-Chao ZHANG ;
Chinese Journal of Orthopaedic Trauma 2004;0(09):-
Objective To investigate effects of volume,distribution and leakage of bone cement in ver- tebroplasty on the adjacent intervertebral bodies using an experimentally calibrated and anatomically accurate fi- nite-element model of elderly L4-L5 vertebral bodies.Methods Computed tomography(CT)scanning was done,at 1 mm intervals,on L4-L5 vertebral bodies of the lumbar spine of an old man cadaver that had no abnormal findings on rnentgenograms.The L4-L5 motion segment data were obtained from the CT scans to establish in computer a three-dimensional finite element model of L4-L5 functional spinal unit(FSU)with the software Ansys 7.0.Compressive load after virtual vertebroplasty on the damaged model,unipedicular and bipedieolar injections and leakage of cement into the intervertebral space,and the resulting endplate and disc stresses of the adjacent vertebral bodies were analyzed in various spatial distributions of the filling material and different loading conditions. An anatomically accurate finite-element model of elderly L4-L5 vertebral bodies was developed.Results The FSU study suggested that changes in stress and strain at adjacent levels were minimal.Furthermore,endplale and disc stresses of the adjacent vertebral bodies were not influenced by bone cement filling volumes or distributions except under bending,whereas asymmetric distribution and leakage of bone cement to the disk increased the stress of adjacent endplates.Conclusion Since asymmetric distribution and significant leakage of cement into the intervertebral space can increase the stresses of endplates of adjacent vertebral bodies and may lead to a fracture, symmetric placement of cement in operation should be achieved and leakage of cement avoided.
8.Clinical Study on Tiaogan Lipi Huaji Prescription Combined with High Intensity Focused Ultrasound for Treatment of Locally Advanced Pancreatic Cancer
Na LI ; Qing ZHANG ; Sheng LIU ; Wenshuo ZHAO
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(7):23-27
Objective To investigate the effects ofTiaogan Lipi Huaji Prescription combined with high intensity focused ultrasound (HIFU) for the treatment of locally advanced pancreatic cancer.Methods Randomized, parallel controlled study was designed to select 60 patients with locally advanced pancreatic cancer, which were divided into treatment group and control group by random number table method, 30 cases in each group. The treatment group was givenTiaogan Lipi Huaji Prescription, one dose per day, twice a day, orally, 28 d as a treatment course, 6 courses in total; at the same time, HIFU was given to the treatment group, 30–60 min each time, once every other day, 2–3 times each week, and the tumor was completely covered for 2 times. The control group was given gemcitabine 1000 mg/m2, intravenous infusion, 1st, 8th d, 21–28 d for a cycle, a total of 6 cycles. The clinical benefit response, QOL-C30 score, survival status, progression-free survival (PFS), CA199 and CEA levels were observed.Results Clinical benefit rates of treatment group and control group were: the treatment group 1, 3, 6 months clinical benefit rate was 64.29%, 63.16%, and 50.00%, respectively; control group clinical benefit rate was 48.15%, 40.00%, and 44.44%, respectively, with statistical significance (P<0.05). The difference of QOL-C30 score between the two groups at the same time wasstatistically significant (P<0.05). During the treatment 1 to 6 months, CA199 in the two groups showed a downward trend, without statistical significance (P>0.05). CEA levels in the treatment group were lower than those in the control group at the same time with statistical significance (P<0.05), except for 2 and 6 months. The survival rates of treatment of 3, 6, and 12 months in the treatment group were 82.14%, 46.42% and 32.14%, respectively, and were 96.67%, 59.32% and 37.04%, respectively in the control group, without statistical significance (P>0.05). The survival time of the treatment group was (253.90±18.06)d in the control group and (246.77±17.31)d in the control group, without statistical significance (P>0.05). PFS was (136.73±16.22)d in the treatment group and (145.77±19.74)d in the control group, without statistical significance (P>0.05).ConclusionTiaogan Lipi Huaji Prescription combined with HIFU for the treatment of locally advanced pancreatic cancer has certain clinical efficacy, which can improve patients' life quality.
9.Treating non-alcoholic fatty liver disease patients of Gan stagnation Pi deficiency syndrome by tiaogan lidi recipe: a randomized controlled clinical trial.
Qiang YU ; Sheng-sheng ZHANG ; Tao ZHOU ; Ying XIONG ; Lu-qing ZHAO ; Yang DING
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(4):401-405
OBJECTIVETo evaluate the efficacy and safety of tiaogan Lipi Recipe (TLR) in treating non-alcoholic fatty liver disease (NAFLD) patients of Gan stagnation Pi deficiency syndrome (GSP-DS).
METHODSA randomized, double blind, placebo-controlled clinical trial was performed. Totally 99 NAFLD patients of GSPDS were randomly allocated into two groups, 66 patients in the treatment group (treated with-TLR, one dose per day) and 33 patients in the control group (treated with placebos, one dose per day). The therapeutic course for all was 12 weeks. All patients received lifestyle interventions including moderate aerobic exercise, moderate caloric restriction, and dietary changes. Clinical symptoms, CT indices, liver functions and blood lipids were observed before and after treatment.
RESULTSAfter 12 weeks of treatment, the total score of clinical symptoms decreased in the two groups (P <0. 01), and it was lower in the treatment group than in the control group (P <0. 05). Liver/spleen CT ratio increased in the treatment group (P <0. 01), and it was higher in the treatment group than in the control group (P <0. 01). After treatment levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT) all decreased in the treatment group (P <0. 05, P <0. 01), while levels of ALT decreased in the control group (P <0. 05). Besides, all the 3 levels mentioned above were lower in the treatment group than in the control group (P <0. 05). Levels of total cholesterol (CHO) and triglyceride (TG) decreased in the two groups (P <0. 05), and they were lower in the treatment group (P <0. 05). Total effective rates of TCM syndrome, abdominal CT, liver functions, and blood lipids were 79. 69% (51/64 cases), 54. 69% (35/64 cases), 67. 65% (23/34 cases), and 67. 39% (31/46 cases) in the treatment group, while they were 56. 25% (18/32 cases), 25. 00% (8/32 cases), 33. 33% (6/18 cases), and 55. 56% (10/18 cases) in the control group. All were superior in the treatment group (P <0.05, P <0.01, respectively).
CONCLUSIONTLR combined with lifestyle intervention could safely and effectively improve clinical symptoms of NAFLD patients of GSPDS, elevate liver/spleen CT ratios, and play a role in liver protection, anti-inflammation, and lowering blood lipids.
Alanine Transaminase ; metabolism ; Aspartate Aminotransferases ; metabolism ; Cholesterol ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Lipids ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Syndrome ; Triglycerides ; gamma-Glutamyltransferase ; metabolism
10.Progresses in the structure and function of Kazal-type proteinase inhibitors.
Qing-Liang ZHENG ; Qing SHENG ; Yao-Zhou ZHANG
Chinese Journal of Biotechnology 2006;22(5):695-700
Proteinase inhibitors are widely distributed in many living organisms and play crucial roles in many biological processes, particularly in regulating the proteinase activity spatially and temporally. However, The Kazal family of serine protease inhibitors is one of the most important and extensively studied protease inhibitor families. This type of protease inhibitor normally consists of one or several domains. Every domain has a highly conserved sequence structure and molecular conformation. It is found that contact residues are hyper variable, which are responsible for the interaction of inhibitors and proteinases. Most of them are in the solvent exposed loop. But P1 residue is the key active site of the interaction between inhibitor and enzyme. The types of the amino acid at P1 site likely play an important role in causing different inhibitory activity. The substitutions at the contact residues cause significant effects on the association constant. By using the Laskowski algorithm, the Ki values of a Kazal domain against six serine proteinases can be predicted from the domain' s sequence alone. At present there are many Kazal proteinase inhibitors found in the organisms, which show important biological functions. This article gives a comprehensive review of the newer developments in the characters and the interaction of the Kazal-type inhibitors.
Amino Acid Sequence
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Models, Molecular
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Molecular Sequence Data
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Protein Structure, Tertiary
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Serine Proteinase Inhibitors
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chemistry
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physiology