1.Fluorescence in situ hybridization detection of chromosome translocations induced by 60Co γ-rays in human lymphocytes dose-response curve and persistence
Xiaoling WANG ; Jin LI ; Zhiquan WANG
Chinese Journal of Radiological Medicine and Protection 2001;21(2):83-86
Objective To study dose-response relationship for chromosome translocation and its persistence measured in human lymphocytes exposed to 60Co γ-rays.Methods The chromosome translocations in human peripheral lymphocytes were detected by fluorescence in situ hybridization using 4# and 7# combination of composite whole chromosome-specific DNA probes and Giemsa stain.Results The dose-response relationship for chromosome translocation induced by 60Co γ-rays in vitro could be described by the function:Y=0.0030+0.0134D+0.0165D2.The frequencies of chromosome translocations induced by 0.5 and 2 Gy γ-irradiation did not diminish over time,so it exhibited excellent persistence.Conclusions The results indicate that retrospective dose-reconstruction can be accomplished using chromosome translocation frequency.
2.Diterpene ginkgolides meglumine injection inhibits oxidative stress induced by oxygen-glucose deprivation by activating Akt/Nrf2 pathway in SH-SY5Y cells
Zhiquan JIN ; Zhiliang XU ; Qiu LIU ; Jun ZHOU ; Liang CAO ; Gang DING ; Zhenzhong WANG ; Wei XIAO
Chinese Journal of Pharmacology and Toxicology 2017;31(1):65-72
OBJECTIVE To investigate the protective effects and mechanism of diterpene ginkgolides meglumine injection (DGMI) against oxidative stress induced by oxygen-glucose deprivation (OGD) in SH-SY5Y cells. METHODS SH-SY5Y cells were divided into five groups: normal control, model control (OGD group) and drug(25 mg · L- 1) administration groups including DGMI group, extract of ginkgo biloba leaves injection group (EGBLI) and lactones ginkgo biloba injection group (LGBI). The cells suffered from oxygen-glucose deprivation (OGD) for 4 h, followed by reoxygenation with drugs for 6 h. Then, cell viabilities were detect using CCK-8 assays, reactive oxygen species (ROS) levels using fluorescence probe DCFH-DA and superoxide dismutase (SOD) activities using WST-1 test. Western blotting was used to detected protein levels of hemeoxygenase-1(HO-1), NAD(P)H, quinone oxidore?ductase l (Nqo1), protein kinase B (Akt), phosphorylated Akt (p-Akt), nuclear factor-E2-related factor2 (Nrf2) and phosphorylated Nrf2 (p-Nrf2). The cells were induced by OGD for 4 h, followed by reoxygen?ation and DGMI for 1 h, combined with different concentrations of PI3K inhibitor (LY294002) (at the final concentration of 12.5, 25 and 50 μmol · L-1) before the protein levels of AKT, p-AKT, Nrf2 and p-Nrf2 were detected by Western blotting. RESULTS SH-SY5Y cells induced by OGD for 4 h resulted in an increase in ROS(P<0.01), but a decrease in cell viabilities(P<0.01), SOD activities(P<0.01), and antioxidant protein levels ( Akt, p-Akt, Nrf2, p-Nrf2, HO-1 and Nqo1) (P<0.01). Compared with OGD group, treatment with reoxygenation and drugs (DGMI,EGBLI and LGBI respectively) for 6 h resulted in a decrease in ROS (P<0.01), but an increase in cell viabilities, SOD activities and antioxidant protein levels of p-Nrf2, HO-1, Nqo1 and p-Akt(P<0.05,P<0.01). DGMI group showed the best efficiently. Moreover, after OGD for 4 h, compared with DGMI group, combining reoxygenation and DGMI with LY294002 for 1 h resulted in a concentration-dependent inhibition of the protein levels of p-AKT and p-Nrf2(P<0.01). CONCLUSION DGMI 25 mg · L-1 can inhibit oxidative stress in SH-SY5Y cells induced by OGD by increasing the activity and expression of Nrf2 through PI3K/Akt pathway, which may be one of the mechanisms by which DGMI protects neurons from stroke.
3.Study on NO Inhibitory Activity of Total Lactones fromAndrographis paniculata
Fangfang XU ; Zhiquan JIN ; Wei SHI ; Yimin MA ; Jun ZHOU ; Zhenzhong WANG ; Yuan BI ; Zhimin WANG ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(5):1061-1065
This study was aimed to explore the suppression of nitric oxide (NO) production in RAW264.7 cells by total lactones fraction from Andrographis paniculata. The inflammatory model in vitro was established by stimulating the RAW264.7 cells with lipopolysaccharide (LPS). The NO production and inhibitory rate were determined by Griess assay. Cytotoxicity was evaluated by MTT method. The results showed that total lactones fraction ofA. paniculata suppressed NO production in a concentration-dependent manner in the concentration range from 5 to 50μmol·L-1 and their IC50 values were 8.58μmol·L-1, 11.52μmol·L-1 and 8.94μmol·L-1, respectively. In this condition, major constituents were andrographolide (5-60μmol·L-1), dehydroandrographolide (5-100μmol·L-1) and neoandrographolide (5-100μmol·L-1) inhibited NO production in a dose-dependent manner with an IC50 values of 17.54μmol·L-1, 49.54μmol·L-1 and 41.80μmol·L-1, respectively. It was concluded that the NO inhibitory activity of total lactones fromA. paniculata was better than three active ingredients, which may be able to provide a theoretical foundation and scientific basis for the preparation and clinical application of total lactones fraction fromA. paniculata.
4.Study on CIX-M type ESBLs-producing Escherichia coli and Klebsiella pneumoiae in Guangzhou
Chao ZHUO ; Danhong SU ; Hongyu LI ; Luxia WANG ; Kang LIAO ; Mei WANG ; Zhiquan ZHI ; Zhonghui GUO ; Yanchao WEI ; Suina GENG ; Guangyao JIN ; Nanshan ZHONG
Chinese Journal of Laboratory Medicine 2009;32(10):1114-1119
Objective To study phylogenies, epidemiology and genetic environment of CTX-M type of ESBLs produced by Escherichia coli and Klebsiella pneumoniae isolated from nine hospitals in Guangzhou. Methods The phylogenies of CTX-M type of ESBLs were analyzed by PCR Genetic environment of CTX-M-15 encoding gene (bla_(CTX-M-15)) were investigated by conjugation test and plasmid analysis. The clonal relationship of strains producing CTX-M-15 was determined by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). Results A total of 361 ESBLs-producing isolates of Escherichia coli and Klebsiella pneumoniae were collected. 67.3% of ESBLs strains were detected to produce CTX-M-type ESBLs, and the commonest genotypes in Escherichia coli and Klebsiella pneumoniae were CTX-M-14 (35.4% and 28.3%), CTX-M-15(21.5% and 26.1%) EBIC-PCR products of all CTX-M-15-producing strains show 39 strains of Escherichia coli were classified into 27 genotypes while 43 strains of Klebsiella pneumoniae were divided into 30 genotypes. Furthermore, the genotypes of CTX-M-55, CTX-M-19, CTX-M-27, with ceftazidime-hydrelyzing activity, were detected in this study. The great majority of bla_(CTX-M-15) genes were found to locate on a 65 000 bp-conjugative plasmid, and there was no blaTEM-1, bla_(OXA-1), blaDSA-1 or aac (6')-Ib-cr gene coexisted on the plasmid, ISEcp1-like insertion sequences, relative to mobilization of bla_(CTX-M-15) gene, were detected in all bla_(CTX-M-15) positive strains, and the distances between the end of ISEcp1-like insertion sequences and the start cedon of bla_(CTX-M-15) were equal, with 48 base pairs. Conclusion CTX-M-14 is still the most common genotype of ESBLs in Guangzhou, but high prevalence of CTX-M-15 ESBLs hydrolyzing ceftazidime already appears in south China.
5.Rapid Identification and Quality Analysis of Olibanum by TGA-DTA Thermal Analysis
Yongheng WEI ; Zhiquan ZHENG ; Juyuan LUO ; Guohang JIN ; Gongsen CHEN ; Yanni LI ; Xinjie LI ; Jinli SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(4):629-635
This study was aimed to quickly identify Chinese medicine Olibanum.Thermal analysis method was used on the quality analysis of Chinese materia medica (CMM).A total of 25 batches of Olibanum on the market were collected.This study examined three important factors of temperature range,heating rate,powder mesh on the TGA and DTA thermal analysis experiments.And a method of rapid authentication of medicinal materials using TGA and DTA feature maps was built.Methods of the first-order points,connection on thermogravimetric analysis and heat enthalpy calculation were adopted in the quantitative analysis of Olibanum.The results showed that the best condition of TGA and DTA experiment on Olibanum was confirmed.The temperature range is 50-750℃.The heating rate is 20℃· min-1.The powder mesh is 100 mesh.Under these conditions,good quality goods of Olibanum,counterfeit Olibanum and adulterants of Olibanum could be distinguished through the characteristic peak (T1=447 ± 5℃,T2=549 ± 5℃,T3=350 ± 5℃),thermogravimetric analysis (TV-max,△W2+△W3) and thermal enthalpy analysis (△H).It was concluded that the TGA-DTA technology was simple.It was thought to be a rapid,accurate and simple new method for Olibanum identification and quality analysis.
6.Assay of eupatilin and arteanoflavone in Artemisia anomala by HPLC
Zhiquan QIU ; Qingqing CAO ; Weifeng TAN ; Jin ZHOU ; Lei LÜ
Journal of Pharmaceutical Practice 2016;34(2):163-166
Objective To determine the concentration of eupatilin and arteanoflavone in A rtemisia anomala by high per-formance liquid chromatography (HPLC).Methods A rtemisia anomala was extracted by ultrasonic for 60 minutes with 10 times volume of methanol.The HPLC was performed on a SHISEIDO MG-C18 column (3.0 mm × 100 mm,3μm).The mobile phase was a mixture of acetonitrile (ACN) and 0.1% formic acid (40:60,V/V ).The detection wavelength was 350 nm,the column temperature was 25 ℃ and the injection volumn was 5μl.Results Eupatilin and arteanoflavone were separated at base-line within 15min with good linearity.The method validation results show that the precisions,repeatability and stability were all in the normal range.The low,medium and high level recoveries of eupatilin were 100.26%,99.58%,102.24%,and those of arteanoflavone were 99.09%,101.12%,101.43%,respectively.Conclusion The method was rapid,simple,reproductive and accurate.It can be used to control the quality of Artemisia anomala.
7.Diterpene ginkgolides meglumine injection attenuates oxygen-glucose deprivation-induced apoptosis of nerve cells via inhibition of calpain signaling pathway
Qiu LIU ; Zhiliang XU ; Zhiquan JIN ; Jun ZHOU ; Yu& ; prime ; an BI ; Zhenzhong WANG ; Wei XIAO
Journal of China Pharmaceutical University 2015;46(6):707-711
To investigate the anti-apoptotic effect of diterpene ginkgolides meglumine injection(DGMI)on SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation(OGD/R), and to explore its mechanisms. After 4 h of OGD, the SH-SY5Y cells were treated with 25 mg/L DGMI for 1 h. The release of lactic dehydrogenase(LDH)was measured by cytotoxicity detection kitplus. Cell apoptosis was detected by caspase-3/7 assays. Cell death was detected by ELISA. The concentration of [Ca2+]i in cytoplasm was measured by Fluo-3 AM and the levels of calpain and cleaved capaease-12 were evaluated by western blot. As we expected, DGMI significantly decreased the release of LDH, the concentration of [Ca2+]i, the protein levels of calpain and cleaved caspase-12. Furthermore, DGMI injection also attenuated the activities of caspase-3/7 and the contents of cytoplasmic histone-associated- DNA-fragments. These data demonstrated that the DGMI injection showed good anti-apoptotic effect in SH-SY5Y cells induced by OGD/R. The mechanisms may be associated with the inhibition of Ca2+/calpain/caspase-12/caspase-3 signaling pathway.
8.Sclerosing polycystic adenosis of the parotid gland: a case report and literature review
CHEN Zengquan ; JIN Tingting ; WANG Yan ; WANG Lin ; MAI Lianxi ; HUANG Zhiquan
Journal of Prevention and Treatment for Stomatological Diseases 2019;27(11):729-732
Objective :
To explore the clinicopathological features, diagnosis, treatment and prognosis of sclerosing polycystic adenosis (SPA) and provide a reference for clinics.
Methods :
A case of sclerosing polycystic adenosis of the parotid glands was retrospectively analyzed, and the relevant literature was reviewed.
Results :
A 57-year-old female patient presented with a tumor, which she had noticed for half a month, on the left side of the lower ear, with occasional paroxysmal numbness and no complaint of other discomfort. Resection of the left submandibular area tumor was performed, and the tumor specimen pathological results showed sclerosing polycystic adenosis of the left parotid gland, with no recurrence after six-months follow-up. Sclerosing polycystic adenosis is rare, occurs in the parotid gland and is characterized by a frequently painless, slow-growing mass of the parotid gland. Imaging examination and fine needle aspiration biopsy can only be used as a reference; the diagnosis must include a pathology examination. Histological manifestations showed that abundant sclerotic collagenous stroma was permeated by ductal and acinar lobules, and cystic dilatation of the duct was accompanied by epithelial hyperplasia and diverse ductal cells. Immunohistochemistry of the ductal and acinar cells showed positive expression of cytokeratin (AE1-3 and CAM5.2) and S100 protein. The ducts filled with hyperplastic and dysplastic epithelium were surrounded by an intact myoepithelial layer that was positive for SMA, p63, and calponin, with a Ki-67 index less than 3%. Treatment comprised mainly surgical resection, with a good prognosis. However, one-third of cases relapse: low-grade malignant tumors may occur, with at least one report of invasive cancer.
Conclusion
Sclerosing polycystic adenosis of the salivary gland is rare and has a good prognosis, but patients may relapse easily after surgery. The diagnosis depends primarily on pathological examination. The main treatment is surgical resection, the prognosis is good, and follow-up should be strengthened after surgery.