1.Research progress of cell transplantation in the treatment of osteochondral lesions of the talus
Hao WANG ; Feng QU ; Shenglong DING ; Songmin SUN ; Mingzhu ZHANG
Chinese Journal of Trauma 2023;39(7):665-672
Osteochondral lesions of the talus (OLT) frequently manifest following ankle joint trauma, causing ankle pain, swelling and impaired mobility, thereby significantly impeding daily activities of the patients. Presently, clinical treatment approaches encompass both conservative management and surgical intervention. Conservative management endeavors to alleviate symptoms, while patients experiencing persistent symptoms resort to surgical intervention. Commonly employed surgical treatments encompass bone marrow stimulation, autologous osteochondral transplantation, and allogeneic osteochondral transplantation. Bone marrow stimulation is employed as a therapeutic approach for the management of smaller OLT, demonstrating favorable short-term effectiveness; however, the long-term prognosis remains uncertain. Autologous osteochondral transplantation is a viable option for larger OLT lesions, albeit it carries the potential of complications at the donor site. Conversely, allogenic osteochondral transplantation exhibits a diminished success rate. In recent times, the utilization of cell transplantation techniques has garnered escalating interest in the treatment of OLT due to their capacity to regenerate cartilage resembling hyaline and their diverse range of cellular origins. The authors reviewed the progress of cell transplantation in the treatment of OLT, providing a reference for the clinical treatment.
2.Prokaryotic expression and functional identification of androgen receptor functional domains
Yuqi Zhang ; Mingzhu Ding ; Jinfeng Wu ; Haixiang Zheng ; Lei Zhang ; Xing Wang
Acta Universitatis Medicinalis Anhui 2023;58(2):219-224
Objective:
To construct the full-length prokaryotic expression plasmid of the wild type of androgen receptor (AR) and the truncated body of four functional domains,and to identify the fusion protein by Western blot and electrophoretic mobility shift assay ( EMSA) .
Methods:
Based on the pGEX-4T-1 vector ,the recombinant plasmids were constructed to express the full-length and functional domains of AR. IPTG was used to induce the expression of the recombinant proteins,which were isolated and purified by glutathione sepharose 4B beads under the optimized condition.The specific protein expression in the bacterial lysate and the purified protein isolated with glutathione sepharose 4B beads was identified by Western blot with AR antibody and GST labeled antibody.The purified protein was incubated with a fluorescent probe of the virus,and the complex was detected by electrophoresis in a non-denaturing gel.
Results :
The prokaryotic recombinant plasmids of full length and three functional domain truncated AR were successfully constructed.The recombinant clones were identified by using bacterial culture as a template,and further verified by double enzyme digestion.It showed that there were identical bands in the same sizes as the inserted fragments.The nucleotide and the amino acid sequences were aligned to the reference sequence in NCBI GenBank.The GST fusion protein,GST-AR-NTD + DBD (96 ku) and GST-AR-NTD (86 ku) were successfully induced and verified. The purified protein could be directly combined with the viral genome DNA.
Conclusion
The prokaryotic expression conditions of truncated AR plasmid from the same gene sequence are different.The purified AR protein can be used to understand the direct interaction mechanism between functional domains of AR and other molecules.
3.Prevalence of hypertension based on three separate visits among primaryand middle school students in Suzhou City
Di HAN ; Mingzhu SHEN ; Bing SHI ; Bo HAI ; Ziyao DING ; Jieyun YIN ; Hui SHEN ; Jia HU
Journal of Preventive Medicine 2022;34(2):109-113
Objective:
To investigate the prevalence of hypertension among primary and middle school students living in Suzhou City, Jiangsu Province, so as to provide insights into comprehensive hypertension control among children and adolescents.
Methods:
Primary and middle school students at ages of 7 to 17 years were recruited for a questionnaire survey in Suzhou City using the stratified cluster random sampling method from September to December, 2020, and the height and body weight were measured. Blood pressure was measured at three separate clinic visits according to the national criteria Reference of Screening for Elevated Blood Pressure among Children and Adolescents Aged 7-18 Years ( WS/T 610-2018 ), and the detection of elevated blood pressure was estimated at three separate visits. In addition, factors affecting elevated blood pressure were identified.
Results:
A total of 3 713 students were enrolled, including 1 924 boys ( 51.82% ) and 1 789 girls ( 48.18% ). The detection of elevated blood pressure was 13.63%, 5.36%, and 3.37% at three separate visits, respectively, and the prevalence of hypertension ( elevated blood pressure at all three visits ) was 3.37%. The detection rates of elevated blood pressure were all higher at three visits ( 16.90%, 8.40%, and 5.26% ) among students at ages of 12 to 17 years than among students at ages of 7 to 11 years ( 9.65%, 1.67%, and 1.07%, P<0.05 ). The detection of elevated blood pressure was significantly higher in boys ( 15.23% ) than in girls (11.91%) at the first visit ( P<0.05 ), while no significant differences were seen at the second or third visit ( P>0.05 ). In addition, higher detection rates of elevated blood pressure were seen in obese ( 27.62%, 11.51%, and 7.06% ) and overweight students ( 17.45%, 6.95%, and 4.85% ) than in students with normal weight ( 9.44%, 3.54%, and 2.15% ) at all three visits, and greater detection rates of elevated blood pressure were found in obese students than in overweight students at the first and second visits ( P<0.017 ).
Conclusions
The prevalence of hypertension was 3.37% based on three separate visits among primary and middle school students in Suzhou City. Measurement of blood pressure at three separate visits within different days is effective to reduce the false positive rate of hypertension and decrease misdiagnosis among children and adolescents.
4.Advances in biodegradation of petroleum hydrocarbons.
Chinese Journal of Biotechnology 2021;37(8):2765-2778
Petroleum hydrocarbon pollutants are difficult to be degraded, and bioremediation has received increasing attention for remediating the hydrocarbon polluted area. This review started by introducing the interphase adaptation and transport process of hydrocarbon by microbes. Subsequently, the advances made in the identification of hydrocarbon-degrading strains and genes as well as elucidation of metabolic pathways and underpinning mechanisms in the biodegradation of typical petroleum hydrocarbon pollutants were summarized. The capability of wild-type hydrocarbon degrading bacteria can be enhanced through genetic engineering and metabolic engineering. With the rapid development of synthetic biology, the bioremediation of hydrocarbon polluted area can be further improved by engineering the metabolic pathways of hydrocarbon-degrading microbes, or through design and construction of synthetic microbial consortia.
Bacteria/genetics*
;
Biodegradation, Environmental
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Hydrocarbons
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Petroleum
;
Petroleum Pollution/analysis*
;
Soil Microbiology
;
Soil Pollutants
5.Clinical application of a prefabricated interocclusal recording cap for making interocclusal records of implant-supported fixed prostheses
Qian DING ; Tingting PU ; Mingzhu HE ; Shimin WANG ; Wenjin LI ; Lei ZHANG ; Jianzhang LIU ; Dai TONG ; Yongsheng ZHOU
Chinese Journal of Stomatology 2021;56(12):1205-1210
Objective:To use a self-developed, prefabricated cap for making interocclusal records in implant-supported fixed prosthetic treatment, and to evaluate its effect of clinical application and accuracy of transferring intra-oral intercuspal position.Methods:Series of prefabricated caps for occlusal recording of implant-supported fixed prostheses were designed based on the healing abutments, and fabricated with three-dimensional (3D) printing. According to the inclusion and exclusion criteria, 12 partially edentulous patients who visited the Department of Prosthodontics, Peking University School and Hospital of Stomatology between November 2020 and September 2021, had lost no fewer than 2 contiguous teeth in distal extension (Kennedy Class Ⅰ or Ⅱ), and had received implant placement 3 months ago were enrolled. Self-control study design was used. Two occlusal records of each included case were obtained by the following 2methods:for the test group, appropriate prefabricated caps were used; and for the control group, polyvinyl siloxane occlusal record was directly set on the healing abutments. The working casts were mounted on the mechanical articulator using the 2 groups of occlusal records successively. Accuracy of occlusal relationship of the mounted casts was evaluated. Diagnostics test was performed to obtain the sensitivity and positive predictive value, which were determined in photographs by comparing the intra-oral occlusal contact points with those in the mounted casts, point-by-point. Virtual casts were taken by intraoral and extraoral scans in intercuspal position and imported to Geomagic Studio 2014. Then the root mean square values of occlusal clearance space between the upper and lower occlusal surfaces of remaining posterior teeth, and the deviations of the interocclusal position of the occlusal surfaces of the remaining teeth in the mandibular arches when superimposing the maxillary arches of the intraoral and extraoral scans were calculated. As a self-control design, paired t test was used. Results:Twelve participants were enrolled, including 3 men and 9 women aged (52.6±12.1) years, and 36 missing teeth. The prefabricated caps could fit on the healing abutments with good retention and stability. The sensitivity of the test group (0.73±0.14) was significantly better than that of control group (0.63±0.12, P<0.01), with no significant difference in positive predictive value between the 2 groups ( P>0.05). The deviations of the interocclusal position of the occlusal surfaces of the remaining teeth were (357.0±140.2) μm for the test group, and (399.4±206.3) μm for the control group, with no significant difference between them ( P>0.05). Conclusions:Interocclusal position record based on prefabricated cap in this study for implant-supported fixed prosthetic treatment can improve the consistency between the intra-oral occlusion and the occlusion in dental casts. This technique has good accuracy, clinical convenience and usability.
6.Biosynthesis of α-lipoic acid in Gluconobacter oxydans increases the production of vitamin C by one-step fermentation.
Yu LIU ; Enxu WANG ; Caihui PAN ; Xiutao DONG ; Mingzhu DING
Chinese Journal of Biotechnology 2019;35(7):1266-1276
In a one-step fermentation system of vitamin C production with Gluconobacter oxydans and Ketogulonicigenium vulgare, a functional module of α-lipoic acid biosynthesis was constructed in G. oxydans. The engineered G. oxydans was co-cultured with K. vulgare to enhance the growth and 2-keto-L-gulonic acid (2-KGA) production of K. vulgare. This one-step fermentation system alleviated the growth inhibition during the mono-culture of K. vulgare and strengthened the interaction between the two bacteria. Moreover, the yield of vitamin C precursor (2-KGA) increased to 73.34 g/L (the control group was 59.09 g/L), and the conversion of D-sorbitol to 2-KGA increased to 86.0%. This study provides a new idea for further optimizing the one-step fermentation system of vitamin C production.
Ascorbic Acid
;
Fermentation
;
Gluconobacter oxydans
;
Rhodobacteraceae
;
Thioctic Acid
;
biosynthesis
7.Effects of hypoxia on adiponectin in tongue squamous cell carcinoma
Xiaohong GUO ; Chong DING ; Junlin ZHANG ; Xiaojing LAI ; Yuan PENG ; Fenghua WU ; Mingzhu YIN ; Yi LI ; Gang ZHAO
Chinese Journal of Clinical Oncology 2019;46(16):815-821
Objective: Adiponectin (APN) is an endogenous cytokine that mediates the development and progression of various tumors through its receptors (AdipoRs). The present study aimed to detect the expression and distribution of APN and its receptors (AdipoR1 and AdipoR2) in tongue squamous cell carcinoma (TSCC). Moreover, we explored whether the locoregional expression of APN was reg-ulated by HIF-1α in the hypoxic microenvironment. Methods: The expression and distribution of APN and its receptors in TSCC tissues were analyzed by immunohistochemical. Lentiviral expression vector for HIF-1α shRNA was constructed and stably transfected in TSCC cells to knock down HIF-1α expression. The mRNA and protein expression levels of APN and its receptors were detected using RT-PCR and Western blot, respectively, after hypoxic treatment. Results: The locoregional expression of APN and AdipoR1, but not AdipoR2, was upregulated at the early stages of T1, T2, and/or N0 stage, respectively, in tumor tissues compared to that in control paracancer-ous tissues (P<0.05 or P<0.01). The expression of APN and AdipoR1, but not AdipoR2, in TSCC cells was up-regulated on hypoxic treat-ment. Moreover, the expression of APN and AdipoR1 was down-regulated after shRNA knockdown of HIF-1α under hypoxia. Conclu-sions: The APN-AdipoR1 signaling pathway was activated and regulated by HIF-1α in the hypoxic environment of TSCC tissues.
8.Fitness analysis between the L-sorbosone dehydrogenase modules and Ketogulonigenium vulgare chassis.
Si CHEN ; Nan JIA ; Mingzhu DING ; Yingjin YUAN
Chinese Journal of Biotechnology 2016;32(9):1224-1232
Ketogulonigenium vulgare is an acid-producing strain in the process of two-step vitamin C fermentation. L-sorbosone dehydrogenase (SNDH) is one of the key enzymes during the biosynthesis of 2-keto-L-gulonic acid (2-KGA), the precursor of vitamin C. However, the catalytic mechanism of SNDH is unclear. According to the whole genome sequencing of K. vulgare, two genes encoding sorbosone dehydrogenases, one derived from the chromosome (named as sndhg) and one from plasmid (named as sndhp), were introduced into an industrial strain K. vulgare. The overexpression of gene sndhg had hardly effect on 2-KGA production, and the overexpression of gene sndhp produced an obvious byproduct in the fermentation broth. Combinational expression of sndhg/sndhp with pqqA (obtaining sndhg-pqqA and sndhp-pqqA modules) in K. vulgare resulted in the similar fermentation phenotype to two previous strains. After serial sub-cultivation of co-cultured Bacillus endophyticus with each engineered K. vulgare for 50 d, the conversion rate of 2-KGA increased by 15.4%, 179%, 0.65% and 125% compared with that of the parental K. vulgare with B. endophyticus. This study shows that adaptive evolution of microbial consortium is an effective strategy to increase the fitness between functional modules and chassis, thus quickly getting better strains for production of 2-KGA.
Aldehyde Oxidoreductases
;
genetics
;
metabolism
;
Ascorbic Acid
;
Bacillus
;
Bacterial Proteins
;
genetics
;
metabolism
;
Coculture Techniques
;
Fermentation
;
Industrial Microbiology
;
Microorganisms, Genetically-Modified
;
Rhodobacteraceae
;
enzymology
;
genetics
;
Sugar Acids
;
metabolism
9.Measurement of lumbar zygapophyseal joints by using spiral CT and its clinical value
Fenglei QIAO ; Bin ZHU ; Jun MA ; Lei BAO ; Ningning DING ; Mingzhu DONG
Journal of Practical Radiology 2015;(10):1664-1667
Objective To study the angle and articular facet curvature of lumbar zygapophyseal joints in adults.Methods The lumbar zygapophyseal joints in 120 healthy subjects without lumbar diseases were detected using spiral CT and mutiple planner re-construction.The angle and articular facet curvature of zygapophyseal joints were measured.The differences in the measured param-eters between male and female or different age groups were compared.Results ①There were no significant differences in the angle of the same lumbar zygapophyseal joints between males and females or different age groups (P >0.05);however,the significant differences in the angle between different lumbar segments were found (P <0.05).②There were no significant differences in articu-lar facet curvature of the same lumbar zygapophyseal joints between males and females or different age groups (P >0.05);however, there were significant differences in the curvature between different lumbar segments (P <0.05).The maximum articular facet cur-vature of L3-L4 was 22.1°±6.0°.Conclusion The angle and articular facet curvature of lumbar zygapophyseal joints varies from different lumbar segments with different contributions for the lumbar stability.
10.Metabolomics analysis of taxadiene producing yeasts.
Huifang YAN ; Mingzhu DING ; Yingjin YUAN
Chinese Journal of Biotechnology 2014;30(2):223-231
In order to study the inherent difference among terpenes producing yeasts from the point of metabolomics, we selected taxadiene producing yeasts as the model system. The changes of cellular metabolites during fermentation log phase of artificial functional yeasts were determined using metabolomics methods. The results represented that compared to W303-1A as a blank control, the metabolites in glycolysis, tricarboxylic acid cycle (TCA) cycle and several amino acids were influenced. And due to the changes of metabolites, the growth of cells was inhibited to a certain extent. Among the metabolites identified, citric acid content in taxadiene producing yeasts changed the most, the decreasing amplitude reached 90% or more. Therefore, citric acid can be a marker metabolite for the future study of artificial functional yeasts. The metabolomics analysis of taxadiene producing yeasts can provide more information in further studies on optimization of terpenes production in heterologous chassis.
Alkenes
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metabolism
;
Amino Acids
;
metabolism
;
Citric Acid
;
analysis
;
Citric Acid Cycle
;
Diterpenes
;
metabolism
;
Fermentation
;
Glycolysis
;
Metabolome
;
Metabolomics
;
Yeasts
;
metabolism


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