1.Effect and mechanism of baicalin on the proliferation and migration of human periodontal ligament stem cells induced by lipopolysaccharide
Qiao WANG ; Bo FENG ; Yongzhi GAO
China Pharmacy 2023;34(10):1216-1222
OBJECTIVE To explore the regulatory effects of baicalin on the proliferation and migration of human periodontal ligament stem cells (hPDLSCs) induced by lipopolysaccharide (LPS) and Janus protein tyrosine kinase 2 (JAK2)/signal transduction and transcription activator 3 (STAT3) signaling pathways. METHODS hPDLSCs were divided into control group, LPS group, different concentration baicalin groups (0.1, 1 and 10 mg/L). ELISA method and CCK-8 assay were used to determine the contents of cell inflammatory factors [interleukin 6 (IL-6), IL-1β and tumor necrosis factor-α (TNF-α)] and cell viability, so as to screen the optimal concentration of baicalin for follow-up pathway validation experiments. The cells were then divided into control group, LPS group, optimal baicalin concentration group and inhibitor group (10 μg/mL LPS+1 mg/L baicalin +3 μmol/L JAK2/STAT3 pathway inhibitor AG490). After treated for 24 h, the proliferation rate of hPDLSCs, apoptosis rate, migration rate, invasion cell number, mRNA and protein expressions of Cyclin D1 and caspase-3, the expression of JAK2/STAT3 pathway-related proteins were all detected. RESULTS According to cell inflammatory factors and cell viability, 1 mg/L was selected as the optimal concentration of baicalin. Compared with control group, cell proliferation rate, migration rate, invasion cell number, Cyclin D1 mRNA and protein expression were significantly decreased in LPS group, while cell apoptosis rate, caspase-3 mRNA and protein expression, p-JAK2 and p-STAT3 protein expression were significantly increased (P<0.05). After treated with 1 mg/L baicalin, the above indexes were reversed significantly (P<0.05). The improvement of above indexes in the inhibitor group was more obvious (P<0.05). CONCLUSIONS Baicalin can promote the proliferation, migration and invasion of LPS-induced hPDLSCs and inhibit their apoptosis and inflammation by blocking the JAK2/STAT3 pathway.
2.Determination of zolpidem tartrate tablets after radiation by UHPLC
Wen ZHANG ; Xinhui HUANG ; Xingrui YANG ; Tingting ZHOU ; Jianyi GAO ; Yongzhi LI
Journal of Pharmaceutical Practice 2022;40(1):62-65
Objective To establish a UHPLC method for the determination of zolpidem tartrate tablets after radiation, and to investigate the effect of different radiation doses on the content of zolpidem tartrate tablets. Methods Ultra high performance liquid chromatography was used. The content of zolpidem tartrate tablets irradiated by γ-ray was determined. Using C18 column, acetonitrile methanol-0.05 mol/L phosphoric acid solution (the pH value as 5.5 with triethylamine) (18∶26∶56) was used as the mobile phase. The flow rate was 0.7 ml/min, and the detection wavelength was 254 nm. Results The method validation showed good linearity in the concentration range of 5-80 μg/ml (r=0.999 6); The average recovery was 98.2%, RSD was 1.72%, and the repeatability was 0.87%. The contents of zolpidem tartrate were 105.1%, 106.4%, 102.7% and 105.4% under 0, 8, 25 and 80 kGy radiation. Conclusion UHPLC has accurate results with short analysis cycle in this study. It is suitable for the determination of zolpidem tartrate tablets after radiation. The content of zolpidem tartrate tablets remained basically unchanged after radiation.
3.Clinical effect of anterolateral minimally invasive plate osteosynthesis in treatment of middle-distal humeral shaft fracture
Huayong ZHENG ; Jie GAO ; Yongzhi GUO ; Xiaowei WANG ; Jianzheng ZHANG ; Shaoguang LI ; Zhi LIU ; Tiansheng SUN
Chinese Journal of Trauma 2021;37(6):549-554
Objective:To investigate the clinical effect of minimally invasive plate osteosynthesis (MIPO) through anterolateral approach in treatment of middle-distal humeral shaft fracture.Methods:A retrospective case series study was conducted to analyze the clinical data of 21 patients with middle-distal humeral shaft fracture admitted to 7th Medical Center of Chinese PLA General Hospital from August 2015 to March 2018, including 12 males and 9 females, aged 18-68 years [(31.3±3.6)years]. All were closed fracture. According to AO classification, the fracture were classified as type A in 6 patients, type B in 10 and type C in 5. All patients were treated with anterolateral minimally invasive plate fixation. The operation time, intraoperative blood loss and hospital stay were recorded. The fracture healing was observed after operation. The visual analogue scale (VAS), University of California at Los Angeles (UCLA) shoulder rating scale, and Mayo elbow performance score (MEPS) were used to evaluate the effectiveness before operation and at 2 weeks, 3 months and 12 months after operation.Results:All patients were followed up for 12-26 months [(18.2±2.4)months]. The operation time was 50-82 minutes [(68.2±19.4)minutes], with intraoperative blood loss of 40-95 ml [(60.2±21.3)ml]. The hospital stay was 6-16 days [(6.8±1.2)days]. There was no iatrogenic vascular or nerve injury during operation. The patients with radial nerve injury before operation were all adventitia contusion. The nerve function returned to normal within 3 months after operation. All fractures were healed within 5-10 months [(5.3±1.2)months]. At 2 weeks, 3 months and 12 months after operation, the VAS [(3.6±0.8)points, (2.1±0.4)points, (1.8±0.3)points] was lower than that before operation [(8.3±1.6)points] ( P<0.05); UCLA shoulder rating scale [(31.2±1.5)points, (33.6±0.8)points, (34.6±0.5)points] was higher than that before operation [(28.4±2.3)points] ( P<0.05); and MEPS [(80.2±3.4)points, (93.4±2.2)points, (96.4±3.5)points] was higher than that before operation [(60.5±4.5)points] ( P<0.05). At the last follow-up, the UCLA shoulder rating scale and MEPS showed excellent results. Conclusion:For middle and lower humeral shaft fracture especially for the fracture line relatively distal to the shaft, MIPO technique through anterolateral approach can attain satisfactory results in terms of pain, range of motion of shoulder and elbow joint, and joint function.
4.Study on Anti-inflammatory Effects and Its Mechanism of Water Extract from Inner Mongolia Medicine Agi
Yan XUE ; Xinyuan LIU ; Hongbo GAO ; Yongzhi XUE
China Pharmacy 2020;31(12):1425-1424
OBJECTIVE:To study the anti-inflammatory effects of water extract from Inner Mongolia medicine Agi (i. e. the aerial part of Artemisia frigida ),and to preliminarily explore its mechanism. METHODS :Totally 60 mice were randomly divided into model control group (water),aspirin group (positive control ,0.4 g/kg),Agi water extract high-dose ,medium-dose and low-dose groups (60.0,30.0,15.0 g/kg,by crude medicine ),with 12 mice in each group. After intragastric administration for consecutive 5 d,the ear swelling of mice was induced by 2,4-dinitrochlorobenzene. Another 60 mice were grouped and admini- stered by the same way ,and the foot swelling of mice was induced by carrageenan. The ear swelling degree and inhibitory rate , paw swelling rate were calculated to evaluate the anti-inflammatory effect of Agi water extract. The contents of TNF-α,IL-1β and PGE2 in plantar inflammatory tissue induced by carrageenan in mice were determined by ELISA. The content of MDA was determined by thiobarbituric acid trace method and NO by nitrate reductase method. RESULTS :Compared with model control group,the degrees of ear swelling of mice were decreased significantly in aspirin group and Agi water extract high-dose , medium-dose,low-dose groups (P<0.05 or P<0.01),and the inhibitory rates of ear swelling were 72.3%,71.1%,52.6%, 36.8%,respectively;the paw swelling rates of mice in aspirin group and Agi high-dose,medium-dose groups were decreased significantly,while those were decreased significantly in Agi water extract low-dose group 0.5 h and 4.0 h after inducing inflammation(P<0.05 or P<0.01);the contents of TNF-α,IL-1β,PGE2,NO and MDA in plantar inflammatory tissue of mice in aspirin group and Agi water extract high-dose ,medium-dose groups were decreased significantly (P<0.05 or P<0.01). CONCLUSIONS:Inner Mongolia medicine Agi water extract has obvious anti-inflammatory effects on ear swelling induced by 2,4-dinitrochlorobenzene and paw swelling induced by carrageenan ,and its mechanism may be related to the inhibition of TNF-α, IL-1β and PGE2 generation,the reduction of NO and MDA excessive release in inflammatory tissues.
5.Changes of behavioral and serum inflammatory factors in rats with acute reserpine-induced comor-bidity of pain and depression
Anna WANG ; Jingjie ZHAO ; Xuesong GAO ; Qian ZHAO ; Li LI ; Yongzhi WANG
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(3):230-234
Objective To investigate the effects of acute reserpine-induced pain and depression co-morbidity model on behavior and related inflammatory cytokines in rats. Methods Twelve male 8-week-old SD rats were randomly divided into control group and model group,6 in each group. Rats in the model group were intraperitoneally injected with reserpine(1 mg/kg/d)for 3 days to establish the model. Rats in the con-trol group were injected with the same amount of distilled water. The pain threshold of rats was measured by Von Frey Hairs and Hot Plate Analgesia Test. The depression behavior of rats was evaluated by open-field test,Forced Swimming Test and Sucrose Consumption Test. Serum IL-1β, IL-6, IL-18 and TNF-α content were detected by ELISA. Results Compared with the control group(1d (15. 00±0. 00) g;3d ( 13. 20±4. 03)g),the PWTs of the model group (1d (6. 20±0. 45) g;3d (4. 20±1. 64) g) decreased significantly(t=44. 00,4. 63,both P<0. 05). Compared with the control group (( 8. 82 ± 1. 08) s),the PWTL (( 3. 16 ± 0. 24)s) was significantly reduced on the first day in model group,and the difference was statistically signifi-cant (t=11. 48,P<0. 05). Compared with the control group (( 1 815. 18± 541. 40) cm,(98. 20± 26. 25) s, (87. 78±9. 38)g),the total distance of the open-field test ((948. 91±494. 35)cm)significantly shortened (t=2. 64,P<0. 05),swimming time ((143. 60±21. 45)s) was significantly prolonged (t=-2. 65,P<0. 05), and the sucrose consumption (( 22. 23 ± 6. 97) g) significantly decreased (t=12. 55,P<0. 05) in model group. Compared with the control group (( 285. 80 ± 11. 93) ng/ml, ( 233. 07 ± 8. 47) ng/ml,( 280. 41 ± 14. 31) ng/ml,( 213. 10 ± 33. 87) ng/ml), serum levels of IL-1β, IL-6, IL-18, TNF-α in model group ((471. 23±24. 15) ng/ml,(364. 82±17. 16) ng/ml,(471. 81± 28. 98) ng/ml,(821. 19±93. 16) ng/ml) increased significantly(F=-15. 39,-15. 39,-13. 24,-13. 72,all P<0. 05). Conclusion A large number of intraperitoneal injections of acute reserpine can cause hyperalgesia,depressive behavior and serum inflamma-tory factors in rats,effectively simulating the symptoms of pain and depression,and can be used as a model of pain and depression comorbidity for biological mechanisms and treatment research.
6.Regulation of Chaihu Shugan Powder on kynurenine in raphe nucleus of depressive rats induced by reserpine
Jingjie ZHAO ; Xuesong GAO ; Anna WANG ; Yongzhi WANG ; Yi DU ; Li LI
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(6):481-486
Objective To investigate the antidepressant effects of Chaihu Shugan Powder(CSP) on depressive rats induced by reserpine and its influences on the kynurenine (KYN),indoleamine 2,3-dioxyge-nase(IDO),interleukin-6(IL-6) and tumor necrosis factor-α( TNF-α). Methods Forty rats with similar behavior results were divide into 4 groups randomly,including Control group(Con),Model group(Res),Flu- oxetine group(Res+Flu) and Chaihu Shugan Powder group(CSP). The depressive rat model was established by intraperitoneal injection reserpine. The rats in Res+Flu group were administered with fluoxetine by intrap-eritoneal and rats in Res+CSP group were administered with CSP by intraperitoneal. After 14 days,the be-havior of rats was measured and then the rats were executed and sampled. The content of tryptophan and kynurenine in raphe nuclei tissue were detected. The mRNA expression level of IDO,IL-6,TNF-α in raphe nuclei tissue were detected. Results ( 1) Compared with Con group (( 81. 81 ± 36. 13) s, ( 83. 51 ± 5. 34)%), the swimming immobility time((150. 50±31. 45)s) in Res group increased(t=68. 7, P<0. 05) and the sucrose perference (59. 73±11. 30)%) in Res group decreased(t=23. 8,P<0. 05). Compared with Res group, the swimming immobility time in Res+Flu group((114. 90± 14. 29) s) and Res+CSP group ((111. 7±11. 34)s) decreased(t=35. 6,35. 8,both P<0. 05). Compared with Res group, the sucrose pref-erence in Res+Flu group((78. 21±10. 07)%) increased(t=18. 3, P<0. 05). (2)Compared with Con group (KYN/TRP:(0. 023±0. 016),IDO mRNA:(1. 00±0. 05),IL-6 mRNA:(1. 00±0. 58),TNF-α mRNA:(1. 00±0. 32)), the activity of IDO(KYN/TRP(0. 039±0. 003)) and the mRNA levels of IDO mRNA(3. 63± 0. 31),IL-6 mRNA(2. 36±0. 23),TNF-α mRNA( 3. 56± 0. 14) of Raphe Nuclei tissue in Res group were significantly increased (t=21. 2,12. 9,38. 3,19. 7,all P<0. 05). Compared with Res group, the activity of IDO(KYN/TRP(0. 030±0. 013)),the mRNA expression levels of IDO mRNA( 1. 56±0. 36),IL-6 mRNA (1. 62±0. 16),TNF-α mRNA(2. 64±0. 20)of Raphe Nuclei tissue in Res+Flu group were significantly de-creased(t=38. 8,15. 8,12. 8,26. 4,all P<0. 05). And compared with Res group,the activity of IDO( KYN/TRP(0. 028±0. 021)) ,the mRNA expression level of IDO mRNA( 1. 33± 0. 29),IL-6 mRNA(1. 36± 0. 34),TNF-α mRNA(1. 93±0. 21)of raphe nuclei tissue in Res+CSP group were also significantly decreased (t=23. 21,17. 3,19. 8,29. 8,all P<0. 05). Compared with Res+Flu group,the level of IDO mRNA and in-flammatory factors' mRNA in Res+CSP group were significantly decreased(t=18. 3,20. 8,31. 5,all P<0. 05). Conclusion Chaihu Shugan Powder has antidepressant effect,and the mechanism is related with de-creasing the inflammatory factors,inhibiting IDO activation and decreasing the IDO mRNA.
7.Hodgkin lymphoma in children: clinical analyses of 20 cases
Yongzhi ZHENG ; Jian LI ; Xueling HUA ; Hao ZHENG ; Zaisheng CHEN ; Ling ZHENG ; Cai CHEN ; Mei LI ; Chunxia CAI ; Jinghui YANG ; Yiqiao CHEN ; Yingying CHEN ; Qinli GAO ; Shaohua LE ; Jianda HU
Journal of Leukemia & Lymphoma 2018;27(10):595-599
Objective To summarize the long-term outcomes and safety of childhood Hodgkin lymphoma (HL) with protocol ABVD. Methods The clinical data of 20 children with HL admitted to the Union Hospital of Fujian Medical University from July 2010 to June 2017 were retrospectively analyzed. Among the 20 children with HL, 15 were male and 5 were female. The median age of initial diagnosis was 6.5 years old (3-12 years old). The pathological types were as follow: 1 case was nodular lymphocyte-predominant HL (NLPHL) and 19 cases were classical HL (cHL), including 9 cases of mixed cell type, 9 cases of nodular sclerosis type and 1 case of lymphocyte rich type. Basing on Ann Arbor staging system, 1 patient was evaluated as stage Ⅰ, 4 patients were stage Ⅱ, 10 patients were stage Ⅲ, and 5 patients were stage Ⅳ. There were 3 patients in the low-risk group, 7 patients in the intermediate-risk group, and 10 patients in the high-risk group. There were 9 patients with B symptoms. All patients were treated with the ABVD regimen. Results All the 20 patients completed all chemotherapy courses. After 2 courses, the effective rate was 100%(20/20), including 12 cases of complete remission (CR) and 8 cases of partial remission (PR). After the treatment, 19 cases achieved CR, and at the end of the 6 courses, the evaluation showed that 1 case had residual lesions. Follow-up to February 2018, clinical symptoms of 18 cases achieved CR, 2 cases relapsed (all high-risk group); the median follow-up time was 42 months (10.1-87.9 months), the overall survival rate was 100 % (20/20), the estimated 5-year rate of freedom from treatment failure (FFTF) was (89.1 ±7.3) %.Conclusions According to the risk stratification, ABVD regimen has good safety and long-term efficacy for children with cHL. Even the patients in low-risk or intermediate-risk group do not achieve CR after 2 courses and do not receive radiotherapy, the prognosis of them is still good.
8. Establishment of experimental method for bone-conducted vibration elicited vestibular-evoked myogenic potentials in healthy guinea pigs
Zichen CHEN ; Juan HU ; Yuzhong ZHANG ; Xinyu WEI ; Yongzhi YANG ; Ying GAO ; Yong XU ; Ying CHENG ; Feiyun CHEN ; Min XU ; Qing ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2018;53(12):939-943
Objective:
To estabilsh animal methods of bone-conducted vibration elicited cervical vestibular-evoked myogenic potentials (BCV-cVEMP) and ocular vestibular-evoked myogenic potentials (BCV-oVEMP) in healthy guinea pigs.
Methods:
Eleven healthy (250-350 g) and awake guinea pigs were selected and undertake conventional BCV-cVEMP and BCV-oVEMP examination in prone position. Parameters of waveforms were cauculated.
Results:
The BCV-cVEMP and BCV-oVEMP both could be elicited in 100% (22/22) in guinea pigs respectively, threshold was (85.5±10.8)dB SPL and (90.7±10.6)dB SPL for cVEMP and oVEMP; n1 latency was (4.5±1.3)ms and (4.3±1.5)ms for cVEMP and oVEMP; p1 latency was (5.8±1.4)ms and (5.6±1.7)ms respectively; n1-p1 interwave latency was (1.2±0.4)ms for cVEMP and (1.4±0.6)ms for oVEMP, amplitude was (21.5±17.3)μV and (24.0±16.3)μV respectively.
Conclusion
Both BCV-cVEMP and BCV-oVEMP can be successfully elicited in healthy guinea pigs.
9.Construction of a recombinant Escherichia coli for high trehalose production.
Chao GAO ; Shan ZHANG ; Yongzhi HE ; Jianzhong HUANG ; Zhiyang DONG
Chinese Journal of Biotechnology 2015;31(12):1784-1788
Trehalose, a compatible solute, is widely used in food, cosmetics, pharmaceutical products and organ transplantation. Nowadays, trehalose is mostly produced by enzymatic synthesis with many secondary products and lowpurity. In this study, high amount of trehalose was produced by recombinant E. ccli fermentation. First, a bifunctional trehalose gene TPSP was amplified from genome of C. hutchinscoii. Second, an expression vector pTac-HisA containing TPSP was constructed and transformed into the host E. coli. Expression of this bifunctional enzyme-TPSP converted glucose to trehalose. The result suggested that TPSP from C. hutchinsonji has been successfully expressed in E. ccoi. High amount of extracellular trehalose generated from glucose by whole-cell catalysis and After optimization, the production of trehalose in shake flasks was improved to 1.2 g/L and the relative conversion rate reached 21%. The production in bioreactor reached 13.3 g/L and the relative conversion rate reached 48.6%. It is the first time to realize the functional expression of the bifunctional enzyme-TPSP of C. hutchinsonii in E. coli and achieved the conversion form glucose to trehalose. This study laid a foundation for industrial large-scale production of trehalose.
Bioreactors
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Catalysis
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Escherichia coli
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genetics
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Glucose
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Glucosyltransferases
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Industrial Microbiology
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Organisms, Genetically Modified
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Trehalose
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biosynthesis
10.Effect of blunting NF kappa B activation on CYP2 E1 in immunological liver injury rats
Jinxue JIA ; Jindong QIN ; Xuefeng LI ; Xiaolin KANG ; Hongbo GAO ; Yongzhi XUE
Chinese Pharmacological Bulletin 2015;(8):1076-1080
Aim To determine the function of nuclear factor-κB ( NF-κB ) in immunological liver injury of rat model and its effect on CYP2E1 expression, content and metabolic activity. Methods The immunological liver injury rat model was prepared by injection of Ba-cillus Calmette Guérin ( BCG,125 mg · kg-1 ) for 14 days. The hepatic tissue injury was revealed by hema-toxylin and eosin ( HE ) method and serum concentra-tion of alanine aminotransferase( ALT) , aspartate ami-notransferase ( AST ) respectively. CYP450 total con-tent in hepatic homogenate was determined by spectro-photography. The expression of CYP2E1 protein was detected by Western blot analysis. The enzyme kinetics of CYP2 E1 probe drug chlorzoxazone was evaluated by high-performance liquid chromatography ( HPLC ) as-say. Results The results showed that BCG-pretreat-ment ( 125 mg · kg-1 ) significantly increased the weight of liver and spleen, serum levels of ALT and AST(P<0. 01) , and decreased CYP2E1 expression, content and metabolic activity ( P <0. 05 ) . Adminis-tration of ammonium pyrrolidine dithiocarbamate (PDTC) (50, 100, 200 mg·kg-1) reversed the a-bove hepatic injury stimulated by BCG in vivo. Moreo-ver, PDTC dose-dependently inhibited the down regu-lation of CYP2 E1 ( P<0. 05 ) . Conclusion Passiva-tion of NF-κB can inhibit the down regulation of CYP2 E1 in liver tissue of immunological liver injury rats;NF-κB may be involved in CYP2 E1 down-regula-tion.

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