1.Progress in synthetic biology of pinocembrin.
Chinese Journal of Biotechnology 2015;31(4):451-460
Pinocembrin, belonging to flavanons, was isolated from various plants. Pinocembrin has a variety of pharmacological activities, such as neuroprotective effect, antimicrobial activity, and antioxidant efficacy. Pinocembrin was approved as class I drugs to its phase II clinical trial by CFDA in 2009, mainly used for the treatment of ischemic stroke. As a promising compound, the manufacturing technologies of pinocembrin, including chemical synthesis, extraction from plant and synthetic biology, have attracted many attentions. Compared with the first two technologies, synthetic biology has many advantages, such as environment-friendly and low-cost. Construction of biosynthetic pathway in microorganism offers promising results for large scale pinocembrin production by fermentation after taking lots of effective strategies. This article reviews some of recent strategies in microorganisms to improve the yield, with focus on the selection of appropriate the key enzyme sources, the supply of precursors and cofactors by microorganisms, the choice of substance and the level of the key enzyme expression.
Biosynthetic Pathways
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Fermentation
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Flavanones
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biosynthesis
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Industrial Microbiology
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Plants
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Synthetic Biology
2.The relationship between HPV pseudovirus-neutralizing antibody titers and antibody titer determined by ELISA method
Jianqiang LEI ; Qiong SHEN ; Gaoxia ZHANG
Chinese Journal of Microbiology and Immunology 2009;29(11):1049-1054
Objective To study the relationship of HPV pseudo-neutralizing titers detected by two different reporter genes: Zoanthus sp. green fluorescent protein (ZsGreen) and secreted alkaline phosphatase (SEAP) , and the relationship between HPV the pseudovirus-neutralizing antibody titer and the antibody titer determined by ELISA method. Methods The plasmids with expression cassettes of the HPV capsid protein L1 and L2 genes after codon optimization and the plasmid with reporter gene (ZsGreen or SEAP) were co-transfected into 293FT cells. The cell lysate supernatants were collected after 48 h culture, then the pseudovirus was purified through POROS column chromatography from the supernatants. After the titer of pseudovirus bulk were measured, HPV-16 and HPV-18 pseudovirus-neutralization assays were carried out for determining the titer of sera collected from immunized mice with HPV candidate vaccine and Gardasil HPV vaccine. Results In statistical analysis, the two reporter gene systems for the detection of the pseudovirus neutralizing antibody titer are highly relevant to each other (Spearman coefficient; r = 0. 760). And their neutralizing antibody titers bear a high degree of correlation with the antibody titer (Spearman coefficient: r= 0.577 and r =0. 741). Conclusion ZsGreen and SEAP pseudovirus neutralizing antibody titers are highly relevant to each other. The neutralizing antibody and the antibody titer are also relevant. These results reveal some mechanism of HPV vaccines to prevent the virus from invading the host cells, and are absolutely useful in the protection efficiency evaluation of the HPV-16 and HPV-18 candidate vaccines.
3.Exploration on the purchasing mode of hospital equipment
Lei ZHA ; Jianrong DAI ; Jianqiang CAI
China Medical Equipment 2016;(2):134-136,137
Objective:Through the discussion on the purchasing mode of hospital equipment in China, fully understand the advantages and disadvantages of various procurement ways, choose reasonable acquisition method, optimize acquisition method, standardize the procurement management objective.Methods: By using the method of analogy, comparison of the four main acquisition methods. By the comparative analysis, the scope of each purchase way was confirmed. Results: Public bidding was better for more than 500000 yuan or the treasury payment items, competitive negotiation approach applies to 50000 yuan-500000 yuan purchase project. Consultation approach can be applied to 50000 yuan purchase project. Information project procurement preferred to methods of government procurement.Conclusion: Reasonable choices of purchasing and proper optimization has significant meaning for equipment purchasing and management in hospital.
4.Wedge-shaped autograft reconstructing deformity of severe varus knee in primary total knee arthroplasty
Lele DONG ; Jianjun FAN ; Lei WEI ; Haitao DONG ; Jianqiang LIAN
The Journal of Practical Medicine 2015;(18):3046-3049
[Abstrct] Objective The deformity of severe varus knee (varus angulation > 15°) can be treated by many ways in primary total knee arthroplasty , but the efficacy and cost are different. The topic of this study provides a new surgery: autograft (wedge grafts) implant and substitute medial tibial bone defects. Methods From Jan. 2006 to Jan. 2014 , a retrospective study was conducted in the First Affiliated Hospital of BaoTou Mendical College. Out of 687 patients, 721 primary knees were operated. Bone defects in proximal tibia (varus angulation >15° ) were encounted in 16 patients in 21 knees in the pattern of non-inclusive slope bone defects in the mean varus degree of 27°. Their IBM was 31 kg/m2. The average age of patients was (63 ± 5) years. The score of clinical knee society score (KSS) before operation was 18 on average. Among of them, the pain score were 12 points, the average range of motion (ROM) were 12 points. the average funtional score were 20 points. During operation,the slope bone defect of the medial tibial plateau was tailored into wedge-shaped ladder bone defect by osteotomy,and then the defect was filled with the resected tibial plateau autograft in which the thickness and shape were matched completely. The high-intensity cortical part of the autograft was placed in the rim to sustain the tibial prosthesis. Three months after operation ,patients started to walk without the help of crutch. Results The average followup period was 5.3 y , We observed that an average postoperative scores of clinical KSS from 42 points to 88 points with the score of functional KSS from 57 points to 92 points. Fusion was observed by trabeculae crossing from the graft to the tibial metaphysis on plain films at least 6 mouths without collapse, stress fracture, untight and bone ununion. Confirmation of vascularization of the graft was observed in bone scans of nuclide. showing increased uptake in the operated area. Inflammatory makers were normal. Conclusion Autograft (wedge grafts) reconstruction of medial tibial bone defects, is of high rate of healing in implant bones, good restoration of the function and low cost.
5.Downregulated Hsa-let-7f contributes to the loss of type II collagen by targeting interleukin-10/STAT3 signaling pathway in degenerative lumbar scoliosis
Lei WANG ; Tianwang LI ; Jianqiang LIU ; Xiaozong LIU ; Zhaoguo WANG ; Yan TIAN ; Yongxing ZHANG ; Wei WANG
Chinese Journal of Tissue Engineering Research 2016;20(15):2225-2232
BACKGROUND:MicroRNAs (miRNAs) play an important role in a variety of diseases. Investigation of miRNA expression profile in degenerative lumbar scoliosis is beneficial for understanding its pathogenesis, providing a novel therapeutic target. Therefore, we tested the hypothesis that miRNAs promote intervertebral disc degeneration through the interleukin-10/STAT3 signaling pathway, a potential regulator of intervertebral disc degeneration.
OBJECTIVE:To compare the differentialy expressed miRNAs in the intervertebral disc tissues from patients with degenerative lumbar scoliosis and normal controls and to identify specific miRNAs in degenerative lumbar scoliosis folowed by functional validation.
METHODS: An initial screening of miRNA expression in nucleus pulposus tissues by miRNA Solexa Sequencing was performed in samples from 10 patients with degenerative lumbar scoliosis and 10 controls, respectively. Subsequently, differentialy expressed miRNAs were validated using qRT-PCR. The level of differentialy expressed miRNAs in degenerative nucleus pulposus tissues was investigated. Then, functional analysis of the miRNAs in regulating type II colagen expression was carried out. Western blot and luciferase reporter assay were used to further confirm the target gene.
RESULTS AND CONCLUSION: We identified 30 miRNAs that were differentialy expressed (16 upregulated and 14 downregulated) in patients with degenerative lumbar scoliosis compared with controls. Folowing qRT-PCR confirmation, Has-let-7f was significantly down-regulated in degenerative nucleus pulposus tissues as compared with controls. Moreover, its level was correlated with the severity of disc degeneration. Overexpression of Has-let-7f promoted type II colagen expression in nucleus pulposus cels. Knockout of interleukin-10 induced effects on nucleus pulposus cels similar to Has-let-7f. Bioinformatics target prediction identified interleukin-10 as a putative target of Has-let-7f. Furthermore, luciferase reporter assays demonstrated that Has-let-7f altered the expression of STAT3 and matrix metaloproteinase-2. These findings indicate that the downregulation of Has-let-7f induces type II colagen loss by directly targeting inleukin-10, thereby resulting in intervertebral disc degeneration and degenerative lumbar scoliosis. Has-let-7f is likely to be a novel therapeutic target for degenerative lumbar scoliosis.
6.MicroRNA-491-5p is involved in the pathogenesis of degenerative lumbar scoliosis by targeting matrix metalloproteinase 9
Lei WANG ; Tianwang LI ; Jianqiang LIU ; Xiaozong LIU ; Zhaoguo WANG ; Yan TIAN ; Yongxing ZHANG ; Wei WANG
Chinese Journal of Tissue Engineering Research 2016;(2):248-253
BACKGROUND:MicroRNAs are widely involved in the regulation of protein expression, and play a critical role in many physiological and pathological processes in the body. But microRNA expression profile in degenerative lumbar scoliosis is rarely reported and understood. OBJECTIVE:To compare the microRNA expression profile in the normal intervertebral disc and degenerative lumbar scoliosis and to identify degenerative lumbar scoliosis-specific microRNAs, folowed by functional validation. METHODS: Total RNA samples were extracted from the nucleus pulposus tissues of 57 patients with degenerative lumbar scoliosis as experimental groups and the normal nucleus pulposus tissues of 42 patients with lumbar fractures as control group. An initial screening of differentialy expressed microRNAs in the nucleus pulposus tissues by microRNA microarray was performed in 10 samples from each group. Subsequently, differentialy expressed microRNAs were validated using real-time quantitative RCR. The level of differentialy expressed microRNAs in the degenerative nucleus pulposus tissues was investigated. Then, the functional analysis of microRNAs in regulating colagen II expression was carried out. Western blot and luciferase reporter assay were also used to detect target genes. RESULTS AND CONCLUSION:We identified 22 microRNAs that were differentialy expressed (17 upregulated and 5 downregulated) in degenerative lumbar scoliosis patients compared with the controls. Folowing real-time quantitative RCR confirmation, miR-491-5p was significantly down-regulated in degenerative nucleus pulposus tissues in comparison with the controls. Moreover, its level was closely correlated with the pathological grading of disc degeneration. Overexpression of miR-491-5p promoted type II colagen expression in nucleus pulposus cels. Bioinformatics target prediction identified matrix metaloproteinase-9 as a putative target of miR-491-5p. Furthermore, luciferase reporter assays demonstrated that miR-491-5p directly targeted matrix metaloproteinase-9 and affected its protein expression in nucleus pulposus cels. These results show that the downregulation of miR-491-5p induces type II colagen loss by directly targeting matrix metaloproteinase-9, thereby resulting in degeneration of the intervertebral disc and degenerative lumbar scoliosis. This study also underscores the potential of miR-491-5p as a novel therapeutic target in degenerative lumbar scoliosis.
7.Applied anatomy study on blood vessels of perisacral promontory
Lei WU ; Yanfen LUO ; Qing WANG ; Jun YANG ; Yuntao LU ; Tao HUANG ; Jianqiang QIN ; Shizhen ZHONG
Chinese Journal of General Surgery 1997;0(06):-
Objective To provide applied anatomic data for relevant operations of blood vessels of perisacral promontory(BVPSP). Methods The composition of BVPSP including origin, course, diameter of the middle sacral vessels, the distance between the sacral promontory and the sacral 1 transverse trunk were observed on 37 adult cadavers. Result The BVPSP is composed of the common and internal iliac vessels, the superior segment of the middle sacral vessels and the sacral 1 transverse trunk. Middle sacral artery comes from abdominal aorta. Middle sacral veins are thin walled without valves. The average diameter of middle sacral artery and vein is 1.02 mm and 2.53 mm respectively. The distance between the sacral 1 transverse trunk and the sacral promontory is 5.75 mm. Conclusion The composition of BVPSP, especially middle sacral veins, plentiful vascular anastomosis are the anatomical basis leading to massive hemorrhage in the relevant operations.
8.Analysis of the phenominon of anticoagulant led to false platelet reducing and its the correcting method
Huiqiang LU ; Xiao WU ; Jianqiang ZHANG ; Xiaoping LI ; Zhongwei CHEN ; Lei ZHOU ; Dongmei TANG
Chinese Journal of Primary Medicine and Pharmacy 2013;20(3):396-398
Objective To study the influence and solution of pseudothrom bocytopenia (PTCP) caused by anticoagulant EDTA-K2.Methods Sysmex-2100 hematology analyzer was used to analyze PTCP-positive patients' blood samples,count the platelet in samples anticoagulant with EDTA-K2,sodium citrate and heparin,and used the manual counting methods.Results Platelets counts in anticoagulant bood by EDTA-K2 was significantly lower than anticoagulant bood by sodium citrate and heparin.Lots of aggregated platelets were appeared in the EDTA-K2 anticoagulant samples stained by Wright-Giemsa,while there was no aggregated platelets observed in sodium citrate anticoagulant samples and heparin anticoagulant samples.Conclusion Platelet aggregation in patients with PTCP causes patelet counts false reducation.The patients with PTCP is determined by measuring the whole blood (no anticoagulant)by automatic hematology analyzer immediately or manual couting method.
9.Influencing factors of self-resolved mechanism of medical disputes and recommendations
Yu LIU ; Liangyu WEI ; Jianqiang CHEN ; Jie CHEN ; Yajie HU ; Lei WANG
Chinese Journal of Hospital Administration 2011;27(4):282-284
The unsatisfactory operation of the existing mechanism for resolving medical disputes by doctors and patients may be contributed to the instability of this mechanism, as lack of regulation constraints results in huge discrepancies of outcomes between identical cases. Specifically, the uncertainty for doctor-patient consultation in the guidelines, evaluation and decision mechanism,consultation process, and compensation calculation. In this consideration, it is recommended to elevate the mechanism stability of this consultation by means of uniform guidelines, third-party evaluation and decision, standardized consultation process, and unified compensation standard.
10.The advance in synthetic biology: towards a microbe-derived paclitaxel intermediates.
Wei WANG ; Yan YANG ; Xiaodong ZHENG ; Shuqiong HUANG ; Lei GUO ; Jianqiang KONG ; Kedi CHENG
Acta Pharmaceutica Sinica 2013;48(2):187-92
The synthetic biology matures to promote the heterologous biosynthesis of the well-known drug paclitaxel that is one of the most important and active chemotherapeutic agents for the first-line clinical treatment of cancer. This review focuses on the construction and regulation of the biosynthetic pathway of paclitaxel intermediates in both Escherichia coli and Saccharomyces cerevisiae. In particular, the review also features the early efforts to design and overproduce taxadiene and the bottleneck of scale fermentation for producing the intermediates.