1.Reinforcement of Rhodobacter sphaeroides cofactor NADPH to increase the production of farnesol.
Man XU ; Xianzhang JIANG ; Jianzhong HUANG ; Feng QI
Chinese Journal of Biotechnology 2020;36(1):90-99
Farnesol (FOH) is produced by dephosphorylation of farnesyl diphosphate (FPP) derived from two universal building blocks, dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate (IPP). In Rhodobacter sphaeroides these building blocks are generated by MEP pathway, however, many of the biosynthetic reactions and biotransformations in the MEP pathway are limited by low availability of NADPH. Improvement of the amount of intracellular NADPH may enhance the synthesis of FOH. In this study, we utilized the strategies of increasing the production of NADPH and decreasing the consumption of NADPH. The expression of glucose 6-phosphate isomerase (pgi) and glutamate dehydrogenase (gdhA) were inhibited by RNA interference, respectively, and overexpression of 6-glucose phosphate dehydrogenase (zwf) and 6-glucose phosphate dehydrogenase (gnd) in the pentose phosphate pathway were carried out. The results showed that the content of NADPH in the recombinant strains increased significantly, the highest FOH production of RSpgii in the RNA interfered strain was 3.91 mg/g, and the FOH production increased to 3.43 mg/g after zwf gene and gnd gene has been overexpressed. In order to obtain strains with higher FOH production, we used RSpgii as the starting strain, and zwf, gnd and co-overexpressed zwf + gnd gene were overexpressed in RSpgii, respectively. The highest FOH production of the strain RSzgpi reached to 4.48 mg/g which was 2.24 times that of the starting strain RS-GY2.
2.A new method to evaluate hip joint damage in ankylosing spondylitis
Zhengyuan HU ; Siliang MAN ; Xiaojian JI ; Ying ZHANG ; Yingpei MA ; Yiwen WANG ; Jian ZHU ; Jiang-Lin ZHANG ; Feng HUANG
Chinese Journal of Rheumatology 2019;23(1):19-24
Objective In order to assess the structure damage of hip joint in ankylosing spondylitis (AS), a new radiograph-based scoring method was developed according to the radiological characteristics of hip involvement in AS, as well referring to prior existing scoring indexes. Methods A new scoring method consti-tuted of erosion, sclerosis and joint space narrowing was developed, pelvis anterior-posterior plain films acquired from patients with AS at baseline and follow-up were collected and assessed by two physicians who were trained in image reading by radiologists. All films were scored independently and blindly. Intra- and inter-reader reliability were assessed by intra-class correlation coefficient (ICC), the feasibility of this new scoring method was assessed by the mean time acquired to score a plain (two hips), its ability to detect the change of structure damage was assessed by the comparison of score differences between baseline and different follow-ups. The date were analyzed by paired-t test or nonparametric tests. Analysis of Variance (ANOVA) or nonparametric tests were utilized for the comparison of means of quantitative variables among the three groups, while Chi-square test for rates of categorical variables. Results No statistically significant differences existed in demographic data and suspected risk factors among the three groups at baseline (P>0.05). Intra-observer reliability was good (0.84 and 0.89), as well as the inter-observer reliability (0.72), the mean time needed to score was (33 ±10) seconds. Score changes were not statistically significant in the groups with follow-up duration of 1-2 and 3-4 years, but in the group of over 5 years, baseline/final scores assessed by the two observers were (6.0±2.7/7.5±3.7) and (5.6±2.1/7.1±3.6), respectively, both changes were statistically significant (t=2.86, Z=-2.99; P<0.01). Conclusion This new method is not only reproducible and easy to operate in clinic practice, but also can tell the changes of hip joint structure damage in the interval of over 5 years, further validation is requeired to demonstrate its discriminability in large populations.
3.Simultaneous determination of six kinds of components in Buyang-Huanwu decoction by UHPLC-MS/MS
Lu WANG ; Ningfeng ZONG ; Man JIANG ; Chuang LIU ; Taiping YONG
International Journal of Traditional Chinese Medicine 2019;41(2):177-181
Objective To develop the UHPLC-MS/MS method for the determination of amygdalin, paeoniflorin, ferulic acid, calycosin glucosidase, quercetin and formononetin in Buyang-Huanwu decoction. Methods Isocratic elution was carried out with mobile phase consisting of methanol- 2 mM ammonium formate. The separation was performed on Agilent ZORBAX SB-C18 maintained at 35 ℃. The flow rate was 200 μl/min, and the injection volume was 2 μl. The mass spectrometer was operated in the positive and negative ionization electrospray (ESI) mode using multiple monitoring (MRM) for analysis of six components. The mass spectrometric conditions were that ion source temperature 400 ℃, dry gas flow 500 L/h, atomization gas flow rate 75.8 Kpa, spray voltage 4000 V, dry gas temperature 400 ℃. Results The amygdalin, paeoniflorin, ferulic acid, calycosin glucosidase, quercetin and formononetin were all analyzed exactly, and the linear ranges were 0.5-32, 0.2-12.8, 0.1-6.4, 0.8-51.2, 0.4-25.6, 0.08-5.12 ng, respectively. The r were 0.9921, 0.9945, 0.9928, 0.9958, 0.9947, 0.9966, respectively. The recoveries of six analytes ranged from 99.21% to 101.44% and the relative standard deviations were all below 2.05%. Conclusions A sensitive, accuracy and suitable UHPLC-MS/MS method has been developed, and the method could be applied for the determination of amygdalin, paeoniflorin, ferulic acid, calycosin glucosidase, quercetin and formononetin in Buyang-Huanwu decoction.
4.An investigation of iodine nutritional status in water-borne high iodine areas in Shandong Province, 2017
Na LIANG ; Xiaoming WANG ; Man ZHANG ; Wen JIANG
Chinese Journal of Endemiology 2019;38(3):203-206
Objective To observe the iodine nutritional status in historical water-borne high iodine areas in Shandong Province,so as to provide a basis for improving the intervention strategy of high-iodinated goiter in water-borne high iodine areas.Methods In 2017,in 38 historical high-iodine counties (cities,districts) in 7 cities of Shandong Province,each county (city,district) was divided 5 regions according to the east,west,south,north,and middle.One township (street) was taken in each regions,and four administrative villages were selected in each township (street).In each administrative village 15 household salt samples were extracted for non-iodine salt detection;detection of iodine content of drinking water,urine iodine content and thyroid volume of children aged 8-12 years were carried out in 19 high iodine counties (cities,districts).The semi-quantitative method was used to detect the iodine level of edible salt of residents;the arsenic cerium catalytic spectrophotometry method(WS/T 107-2006) was used to detect the water iodine and urinary iodine content;the B-ultrasound method was used to examine the thyroid volume of children.Results A total of 9 703 edible salt samples were collected from residents,including 9 575 non-iodine salt and the consumption rate of non-iodine salt was 98.68%.Among the 38 high iodine counties (cities,districts),20 counties (cities,districts) all stopped supplying iodized salt and 18 counties(cities,districts) partial stopped,the consumption rates of non-iodine salt were 99.37% (6 013/6 051) and 98.54% (3 562/ 3 652),respectively.The iodine content of drinking water in 19 monitoring sites was detected.The median water iodine was 75.50 μg/L,ranging from 8.50 to 420.70 μg/L.Among them,the median water iodine of 9 monitoring sites was > 100 μg/L,still belonged to the water-borne high iodine area.Totally 2 009 children were detected urinary iodine,the median urinary iodine concentration was 256.35 μg/L;the goiter rate of children aged 8-12 years was 7.86% (158/2 009),there was a statistically significant difference in the rate of goiter between children of different ages (x2 =43.649,P < 0.05).Conclusions Although the consumption rates of non-iodine salt in historical water-borne high iodine areas of Shandong Province are at a higher level,the water iodine content of drinking water has a larger span,iodine nutrition in children is over the appropriate level.The coverage of projects to improve water quality and reduce iodine in high iodine areas should be further expanded,and monitoring of iodine nutritional status of key populations should be strengthened.
5.Effect of hydrogen on expression of matrix metalloproteinase-9 in brain tissues of mice with sepsis-associated encephalopathy
Yi JIANG ; Yang YU ; Yaoqi WANG ; Man YANG ; Chao QIN ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2019;39(5):606-609
Objective To evaluate the effect of hydrogen on the expression of matrix metalloproteinase-9 (MMP-9) in brain tissues of mice with sepsis-associated encephalopathy (SAE).Methods A total of 212 clean-grade healthy male C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =53 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H2),group SAE and SAE plus hydrogen group (group SAE+ H2).The sepsis model was established by intraperitoneally injecting human fecal suspension.Sham+H2 and SAE+H2 groups inhaled 2% hydrogen for 1 h at 1 and 6 h after establishing the model,respectively.Postoperative 14-day survival rate was recorded.Evans blue (EB) was injected into the tail vein at 24 h after establishing the model,and the content of EB in brain tissues was calculated.Brain tissues were taken for determination of brain water content.The expression of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) in hippocampal tissues was detected by enzymelinked immunosorbent assay.The expression of MMP-9 and ZO-1 protein and mRNA in hippocampal tissues was determined by Western blot and real-time polymerase chain reaction,respectively,at 6,12 and 24 h after establishing the model.Results Compared with Sham group,the 14-day survival rate was significantly decreased,the EB content in brain tissues,brain water content,and contents of TNF-α,IL-6 and HMGB1 were significantly increased,the expression of MMP-9 protein and mRNA was up-regulated,and the expression of ZO-1 protein and mRNA was down-regulated in SAE and SAE+H2 groups (P<0.05),and no significant change was found in the parameters mentioned above in group Sham+H2 (P>0.05).Compared with group SAE,the 14-day survival rate was significantly increased,the EB content in brain tissues,brain water content,and contents of TNF-α,IL-6 and HMGB1 were significantly decreased,the expression of MMP-9 protein and mRNA was down-regulated,and the expression of ZO-1 protein and mRNA was up-regulated in group SAE+H2 (P<0.05).Conclusion The mechanism by which hydrogen attenuates the blood-brain barrier damage may be related to inhibiting MMP-9 expression and to reducing inflammatory responses in brain tissues of SAE mice.
6.Advances in DNA test for non-neoplastic Epstein-Barr virus associated diseases in children
International Journal of Pediatrics 2019;46(7):508-511
Epstein-Barr ( EB) virus is wildly spreaded in human and associated with many diseases. EBV-DNA load measurement by PCR has been shown to play an important role in the diagnosis of these disea-ses, a method to monitor patients and a prognosis factor of the outcome in many EBV-associated diseases. This review focuses on the development of the EBV-DNA detection technique and the advances of clinical use in sev-eral non-neoplastic EBV-associated diseases in children.
7.An analysis of iodine nutritional status among key populations in non-iodine excess areas of Shandong Province in 2017
Man ZHANG ; Xiaoming WANG ; Na LIANG ; Ru CUI ; Yu WANG ; Wen JIANG
Chinese Journal of Endemiology 2019;38(7):547-549
Objective To analyze the iodine nutritional status among key populations in non-iodine excess areas of Shandong Province in 2017. Methods Totally 81 county-level monitoring sites were selected in 17 cities of Shandong Province. Each county was divided into five areas including the east, west, south, north and the middle according to the location. A township (street) was randomly selected in each area and a primary school was selected from each township (street). Totally 40 students aged 8-10 (half males and half females) were selected in each school for examination of thyroid by B ultrasonography and determination of urinary iodine and salt iodine levels. At the same time, 20 pregnant women were selected from each township (street) for determination of urinary iodine and salt iodine levels. Results A total of 16341 children and 8524 pregnant women urine samples were tested and the medians of urinary iodine of children and pregnant women were 187.7 and 145.4 μg/L, respectively. Totally 16341 children were examined thyroid and the rate of goiter was 2.36% (386/16341). Totally 24170 edible salt samples were tested. The coverage rate of iodized salt was 88.93%(21495/24170), the qualified rate of iodized salt was 90.08%(19363/21495) and the consumption rate of qualified iodized salt was 80.11%(19363/24170). Conclusion Both children and pregnant women in non-iodine excess areas of Shandong Province are in a suitable state of iodine nutrition.
8.In Vitro and In Vivo Evaluation of Antitumor Activity of Ligustrum robustum, A Chinese Herbal Tea.
Hao-Jiang ZUO ; Shan LIU ; Chun YAN ; Li-Man LI ; Xiao-Fang PEI
Chinese journal of integrative medicine 2019;25(6):425-430
OBJECTIVE:
To examine the effect of the aqueous extract of Ligustrum robustum on tumor growth in vitro and in vivo and explore the possible molecular mechanisms.
METHODS:
In in vitro study, cell viabilities of human cervical carcinoma cells (HeLa), human breast cancer cells (MCF-7), human prostate cancer cells (PC-3), human hepatoma cells (7721) and human colon carcinoma cells (SW480) were evaluated with cell counting kit-8. For L. robustum-treated Hela cells, early or late apoptosis were evaluated by annexin V/PI staining. Mitochondrial membrane potential was measured by staining cells with JC-1. Apoptosis was monitored by nuclear morphology based on chromatin condensation and fragmentation by 4',6-diamidino-2-phenylinole (DAPI) staining. Caspase-3 and -8 activity levels were measured by a colorimetric assay. In vivo, to evaluate the possible mechanism of L. robustum-mediated antitumor effect, nude mouse xenograft study was also conducted.
RESULTS:
In in vitro study, L. robustum was found to be toxic to HeLa, MCF-7, PC-3, 7721, SW480, with an half maximal inhibitory concentration value of 2-5 mg/mL (P<0.05). Moreover, externalization of phosphatidylserine, loss of mitochondrial membrane potential, DNA fragmentation and activation of caspase-3 and -8 were detected in L. robustum-treated Hela cells. Using a nude mouse model bearing Hela xenografts, we found that L. robustum reduced tumor volume and tumor weight (P<0.05), but had no effect on body weight and histological damage of important organs. Intraperitoneal injection of L. robustum caused a significant reduction in serum aspartate transaminase and alanine transaminase levels (P<0.05). Furthermore, cleaved caspase-3-positive and terminal nucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells were observed in L. robustum-treated tumor tissues.
CONCLUSIONS
L. robustum inhibits tumor cell growth both in vitro and in vivo by inducing apoptosis in a caspase-dependent way without apparent hepatic toxicity and histological damage, which may offer partial scientific support for the ethnopharmacological claims of L. robustum as a herbal tea for its antitumor activity.
9.Recent progress on obesity-induced myocardial remodeling and its possible mechanism of mitochondrial dyshomeostasis.
Qing WU ; Run-Qing XUE ; Man XU ; Yi LU ; Xiao-Jiang YU ; Long-Zhu LIU ; Wei-Jin ZANG
Acta Physiologica Sinica 2019;71(2):216-224
Obesity is an important risk factor for cardiovascular diseases, which can lead to a variety of cardiovascular diseases including myocardial remodeling. Obesity may induce myocardial dysfunction by affecting hemodynamics, inducing autonomic imbalance, adipose tissue dysfunction, and mitochondrial dyshomeostasis. The key necessary biochemical functions for metabolic homeostasis are performed in mitochondria, and mitochondrial homeostasis is considered as one of the key determinants for cell viability. Mitochondrial homeostasis is regulated by dynamic regulation of mitochondrial fission and fusion, as well as mitochondrial cristae remodeling, biogenesis, autophagy, and oxidative stress. The mitochondrial fission-fusion and morphological changes of mitochondrial cristae maintain the integrity of the mitochondrial structure. The mitochondria maintain a "healthy" state by balancing biogenesis and autophagy, while reactive oxygen species can act as signaling molecules to regulate intracellular signaling. The excessive accumulation of lipids and lipid metabolism disorder in obesity leads to mitochondrial dyshomeostasis, which activate the apoptotic cascade and lead to myocardial remodeling. In this review, we provide an overview of the recent research progress on obesity-induced myocardial remodeling and its possible mechanism of mitochondrial dyshomeostasis.
Humans
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Mitochondria
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pathology
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Mitochondrial Dynamics
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Myocardium
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pathology
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Obesity
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physiopathology
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Reactive Oxygen Species
10.The prediction of baicalin content in the extraction process of Scutellaria baicalensis by near-infrared spectroscopy combined with different variable selection methods
Xue-song LIU ; Si-yu ZHANG ; Man-qian ZHAO ; Jun WANG ; Ye-rui LI ; Jun DAI ; Chuan-zhen TENG ; Xiao KE ; Yong CHEN ; Yong-jiang WU
Acta Pharmaceutica Sinica 2019;54(1):138-143
Near-infrared spectroscopy (NIRS) combined with chemometrics can achieve rapid detection in process analysis. After variable selection, the redundant information is effectively removed and the characteristic variables related to the response values are selected. Compared with global model, the complexity is significantly reduced and the prediction accuracy is also improved. In this study, near-infrared spectroscopy analysis combined with different variable selection methods was applied to achieve the rapid detection of baicalin in the extraction process of

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