1.DNA repair of CHL cells and HeLa cells after DNA damage induced by different oxidative agents.
Ming-zheng LI ; Zhong-chu JIN ; Wei-ya CHEN ; Hong-juan LI
Journal of Zhejiang University. Medical sciences 2004;33(3):235-238
OBJECTIVETo investigate DNA repair in CHL cells and HeLa cells after DNA damage induced by different oxidative agents.
METHODSCHL cells and HeLa cells were exposed to various damaging agents, CHL cells: H(2)O(2) for 25 min, K(2)Cr(2)O(7) for 105 min, doxorubicin (Dox) for 75 min HeLa cells: H(2)O(2) for 25 min, K(2)Cr(2)O(7) for 105 min; then cells were continuously cultured for 0-3 h after washing. Alkaline single cell gel electrophoresis (ASCGE) assay was used to detect DNA strand breaks.
RESULT(1) DNA strand breaks were induced in CHL cells after exposure to H(2)O(2) K(2)Cr(2)O(7) or Dox, which were repaired evidently after continuous culture for 1 h(P<0.01). The damages induced by H(2)O(2) or K(2)Cr(2)O(7) were repaired completely after culture for 2-3 h. However, the demage induced by Dox was repaired incompletely. (2) DNA strand breaks were induced also in HeLa cells after exposure to H(2)O(2) or K(2)Cr(2)O(7), which were repaired evidently after continuous culture for 0.5 h(P<0.01),and completely after culture for 1 h. (3) The regression coefficient related to the rate of comet cells and repair time was statistically different (P<0.05) between CHL cells and HeLa cells.
CONCLUSIONDNA damage induced by Dox is repaired more difficult than that induced by H(2)O(2) or K(2)Cr(2)O(7). The repair initiates immediately after DNA damage in both of cells, but more rapidly in HeLa cells than in CHL cells.
DNA ; metabolism ; DNA Damage ; DNA Repair ; HeLa Cells ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidation-Reduction ; Regression Analysis
2.Mechanism of PC12 cell differentiation induced by total salvianolic acid
Yang SHEN ; Qi ZHANG ; Jia-Qi ZHAO ; Ya-Juan TIAN ; Qin-Qing LI ; Shi-Feng CHU ; Wen-Bin HE
Chinese Journal of Pharmacology and Toxicology 2018;32(2):99-104
OBJECTIVE To study the differentiation of PC12 cells induced by total salvianolic acid (Tsa) and the mechanism. METHODS MTT assay was used to detect the effect of Tsa 0.01, 0.1 and 1.0 μg·L-1on proliferation of PC12 cells and on the cells damaged by oxygen and glucose deprivation (OGD).The number of projections of PC12 cells was statistically analyzed.Western blotting was applied to detect the levels of microtubule-associated protein2 (MAP-2), extracellular signal-regulated kinase1/2 (ERK1/2), phosphorylated ERK1/2(p-ERK1/2), mitogen-activated protein kinase kinase1/2(MEK1/2) and p-MEK1/2 proteins.MEK inhibitor U0126 was examined for its effect on expressions of p-ERK1/2 and ERK1/2 protein in PC12 cells induced by Tsa 1.0 μg·L-1.RESULTS Compared with normal control group, Tsa 1.0 μg·L-1could promote PC12 cell proliferation, and the survival rate was increased by 90%, but the survival rate of PC12 cells was not affected by Tsa 0.01 or 0.1 μg·L-1. Compared with OGD injured group,PC12 cells injured by OGD could be repaired by Tsa 0.1 or 1.0 μg·L-1,and the survival rate was increased to (47.7±1.8)% and (63.2±13.0)%, respectively (P<0.05, P<0.01). Compared with normal control group,Tsa 0.01,0.1 and 1.0 μg·L-1could promote the growth of PC12 cell projections (P<0.01). Western blotting results showed that Tsa could promote the expressions of MAP-2, p-ERK1/2 and p-MEK1/2 proteins, and this effect could be blocked by U0126 inhibitor (P<0.01). CONCLUSION Tsa can induce the proliferation and differentiation of PC12 cells, the mechanism of which is possibly the activation of p-MEK and p-ERK1/2.
3.Clinical and gene mutation features of cystic fibrosis: an analysis of 8 cases.
Na ZHANG ; Jian-Hua LIU ; Ya-Juan CHU ; Jin-Feng SHUAI ; Kun-Ling HUANG
Chinese Journal of Contemporary Pediatrics 2022;24(7):771-777
OBJECTIVES:
To study the clinical features and gene mutation sites of children with cystic fibrosis (CF), in order to improve the understanding of CF to reduce misdiagnosis and missed diagnosis.
METHODS:
A retrospective analysis was performed on the medical records of 8 children with CF who were diagnosed in Hebei Children's Hospital from 2018 to 2021.
RESULTS:
Among the 8 children with CF, there were 5 boys and 3 girls, with an age of 3-48 months (median 8 months) at diagnosis, and the age of onset ranged from 0 to 24 months (median 2.5 months). Clinical manifestations included recurrent respiratory infection in 7 children, sinusitis in 3 children, bronchiectasis in 4 children, diarrhea in 8 children, fatty diarrhea in 3 children, suspected pancreatic insufficiency in 6 children, pancreatic cystic fibrosis in 1 child, malnutrition in 5 children, and pseudo-Bartter syndrome in 4 children. The most common respiratory pathogens were Pseudomonas aeruginosa (4 children). A total of 16 mutation sites were identified by high-throughput sequencing, multiplex ligation-dependent probe amplification, and Sanger sequencing, including 5 frameshift mutations, 4 nonsense mutations, 4 missense mutations, 2 exon deletions, and 1 splice mutation. CFTR mutations were found in all 8 children. p.G970D was the most common mutation (3 children), and F508del mutation was observed in one child. Four novel mutations were noted: deletion exon15, c.3796_3797dupGA(p.I1267Kfs*12), c.2328dupA(p.V777Sfs*2), and c.2950G>A(p.D984N).
CONCLUSIONS
p.G970D is the most common mutation type in children with CF. CF should be considered for children who have recurrent respiratory infection or test positive for Pseudomonas aeruginosa, with or without digestive manifestations or pseudo-Bartter syndrome.
Bartter Syndrome
;
Child, Preschool
;
Cystic Fibrosis/genetics*
;
Cystic Fibrosis Transmembrane Conductance Regulator/genetics*
;
Diarrhea
;
Female
;
Humans
;
Infant
;
Infant, Newborn
;
Male
;
Mutation
;
Respiratory Tract Infections
;
Retrospective Studies
4.Effect of dynamic adjustment of the insertion depth on the cleansing enema
Yan CHU ; Li-Juan JIN ; Ya-Lan KONG ; Yi HUANG ; Zhong-Qing WEI
Chinese Journal of Modern Nursing 2012;18(36):4434-4436
Objective To improve the effect of dynamic adjustment of the insertion depth on the Cleansing enema so as to reduce the suffering of the patients.Methods Totals of 100 patients who required preoperative cleansing enema were selected in this study.The patients were randomly divided into the experimental group (n =50) and control group (n =50).In experimental group,they received the dynamic adjustment of intubation depth,the intubation depth was 7-10 cm at first,then was changed as 20-25 cm,at last the depth was 7-10 cm.In control group,traditional method(the depth was 7-10 cm) was adopted.The number of enema,the amount of enema,the intestinal cleanliness and the degree of satisfaction of two groups were compared.Results Compared with the control group,in the experimental group,the times of enema was decreased significantly [(3.44 ± 1.46) vs(8.78 ± 1.85),t =16.022,P <0.01],the amount of enema used in each case was increased significantly [(797.0 ± 141.0) ml vs (582.0 ± 145.7) ml ; t =7.498,P < 0.01],the intestinal cleanliness(98.0% vs 64.0%)and the degree of satisfaction (92.0% vs 52.0%) were improved obviously (x2 =18.78,19.841,respectively; P < 0.01).Conclusions This cleansing enema discussed in this study which appropriately changes the insertion depth into the anal canal is obviously better than that of traditional method.And such a new method of cleaning enema is worthy of clinical application.
5.Effects of Shuxuening Injection on Lac, Presepsin and NOS levels in sepsis patients
jing Ya LIU ; ming Chun LI ; shan Hong KANG ; rong Jin WANG ; juan Li AN ; na Hong QI ; guo Yong CHU
Chinese Traditional Patent Medicine 2017;39(9):1795-1799
AIM To investigate the effects of Shuxuening Injection (Ginkgo biloba leaf extract) on serum lactic acid (Lac),soluble CD14-st (Presepsin) and nitric oxide synthase (NOS) levels in sepsis patients.METHODS One hundred and eight patients with sepsis treated by routine treatment in our hospital from Jan.2014 to Oct.2016 were randomly divided into two groups,control group and Shuxuening group (therapy group).Two weeks were one therapeutic course.Before the treatment (the onset of patients within 3 hours),at 6 hours and 5 days after the treatment,Lac and Presepsin levels were detected,and the changes of nitric oxide (NO),NOS,inducible nitric oxide synthase (iNOS) and sequential organ failure assessment (SOFA) score were observed.Incidence of major adverse cardiac events (MACE) and 28-day survival were recorded at the same time.RESULTS Before the treatment,there were no significant differences in SOFA score and the levels of Lac,Presepsin,NO,NOS and iNOS between the two groups (P > 0.05).Six hours after the treatment,the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P < 0.05),both the two groups had lower levels of Lac and Presepsin than those before the treatment (P < 0.05);five days after the treatment,the levels of Lac and Presepsin in the two groups were lower than those at 6 hours after the treatment (P < 0.05),the levels of Lac and Presepsin in the therapy group were lower than those in the control group (P < 0.05).The SOFA score,NO,NOS and iNOS levels after the treatment in the therapy group were lower than those in the control group (P < 0.05).The levels of Lac and Presepsin in sepsis patients were positively correlated with SOFA score (r =0.245,0.261,P =0.011,0.006).The patients in the therapy group had lower incidence of MACE and 28-day mortality rate than those in the control group (P < 0.05).CONCLUSION The therapeutic effect of Shuxuening Injection combined with routine treatment on sepsis patients is superior to that of routine treatment,which can improve the prognosis of patients to a certain extent.
6.Effects of electromagnetic pulse on blood-brain barrier permeability and tight junction proteins in rats.
Lian-bo QIU ; Gui-rong DING ; Ya-mei ZHANG ; Yan ZHOU ; Xiao-wu WANG ; Kang-chu LI ; Sheng-long XU ; Juan TAN ; Jia-xing ZHOU ; Guo-zhen GUO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2009;27(9):539-543
OBJECTIVETo study the effect of electromagnetic pulse (EMP) on the permeability of blood-brain barrier, tight junction (TJ)-associated protein expression and localization in rats.
METHODS66 male SD rats, weighing (200 approximately 250) g, were sham or whole-body exposed to EMP at 200 kV/m for 200 pulses. The repetition rate was 1 Hz. The permeability of the blood-brain barrier in rats was assessed by albumin immunohistochemistry. The expression of typical tight junction protein ZO-1 and occludin in both cerebral cortex homogenate and cerebral cortex microvessel homogenate was analyzed by the Western blotting and the distribution of ZO-1 and occludin was examined by immunofluorescence microscopy.
RESULTSIn the sham exposure rats, no brain capillaries showed albumin leakage, at 0.5 h after 200 kV/m EMP exposure for 200 pulses; a few brain capillaries with extravasated serum albumin was found, with the time extended, the number of brain capillaries with extravasated serum albumin increased, and reached the peak at 3 h, then began to recover at 6 h. In addition, no change in the distribution of the occludin was found after EMP exposure. Total occludin expression had no significant change compared with the control. However, the expression level of ZO-1 significantly decreased at 1 h and 3 h after EMP exposure in both cerebral cortex homogenate and cerebral cortex microvessel homogenate. Furthermore, immunofluorescence studies also showed alterations in ZO-1 protein localization in cerebral cortex microvessel.
CONCLUSIONThe EMP exposure (200 kV/m, 200 pulses) could increase blood-brain barrier permeability in rat, and this change is associated with specific alterations in tight junction protein ZO-1.
Animals ; Blood-Brain Barrier ; radiation effects ; Brain ; metabolism ; Capillary Permeability ; radiation effects ; Electromagnetic Fields ; adverse effects ; Male ; Membrane Proteins ; metabolism ; Phosphoproteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Zonula Occludens-1 Protein
7. Effect of Morinda officinalis on serum metabolic spectrum in ovariectomized osteoporosis rats
Wen-Jing LIU ; Juan YANG ; Chu-Tian ZHANG ; Pan SUN ; Mei-Ya HUANG ; Yun-Mei HUANG ; Mei-Ya HUANG ; Yun-Mei HUANG
Chinese Pharmacological Bulletin 2022;38(3):446-453
Aim To study the effect of Morinda officinalis on serum metabolisms in ovariectomized osteoporosis rats based on metabonomics, and explore the mechanism of Morinda officinalis in the treatment of postmenopausal osteoporosis.Methods Forty-eight SD rats were randomly divided into sham operation group, model group and Morinda officinalis group.The Morinda officinalis group was given Morinda officinalis water extract by gavage.The model group and sham operation group were given normal saline by gavage.The bone mineral density(BMD)of the right femur was measured by dual energy X-ray absorptiometry; the maximum load of the tibia bending at three points and the lumbar compression was measured by universal material testing machine.The endogenous metabolites of ovariectomized osteoporosis rats were identified by serum metabonomics, and the potential differential metabolites were screened and identified..Results The BMD and maximum load of the model group decreased significantly, while the Morinda officinalis group increased significantly compared with the model group.The serum metabolic spectrum of the sham operation group was completely separated from that of the model group, and the Morinda officinalis group was close to the sham operation group, suggesting that the body had a tendency to return to normal after intervention of Morinda officinalis.28 metabolites and 5 metabolic pathways were identified to be related to ovariectomized osteoporosis.Morinda officinalis could regulate the contents of stearic acid, uracil and other metabolites, which were related to the biosynthesis of unsaturated fatty acids, the metabolism of pyrimidine and so on.Conclusion Morinda officinalis can prevent ovariectomized osteoporosis by regulating the lipid metabolism and nucleic acid metabolism.
8. HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on Dendrobium officinale and Characteristics Analysis of Different Provenances
Zhi-yun LIANG ; Zhen-shan XIE ; Yue-chun HUANG ; Yuan YUAN ; Chu-juan ZHOU ; Ya-wen WANG ; Gang WEI
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(1):22-28
Objective: To optimize the HPLC characteristic spectrum of flavonoid glycosides of Dendrobium officinale,and identify the common and specific components of different provenances. Method: Kromasil 100-5 C18 column was adopted, with tetrahydrofuran-acetonitrile-methanol (10:22:5)-0.05% phosphoric acid as mobile phase (gradient elution). The detection wavelength was 340 nm,the column temperature was 30℃,and the flow rate was 1.0 mL ·min-1. Result: 13 flavonoid characteristic peaks were marked in 27 batches of D. officinale,and 7 characteristic peaks of 6 flavonoid C-glycosides (vicenin Ⅱ,vicenin Ⅰ,schaftoside,isoschaftoside,violanthin and isoviolanthin) and one flavonoid O-glycosides (rutin) was identified. 7-11 characteristic peaks were detected in different batches of samples. Among them,vicenin Ⅱ was a relatively stable common peak in different source samples,and the characteristic peaks of rutin,schaftoside and isoschaftoside were quite different. According to the relative abundance of the characteristic peaks,the samples could be divided into three categories. Among them,the first category had 10 batches of samples,which mainly came from Danxia landforms of Guangdong,Jiangxi,Fujian and Zhejiang (Wuyi) Province (which called "Danxia landform species") and characterized by detection of obvious peak of rutin. The second category had 11 batches of samples,which mainly came from Yunnan and Guangxi Province (which included "Yunnan Guangnan species" and "Guangxi Tiepilan species") and characterized by detection of violanthin and isoviolanthin. And the third category had 6 batches of samples, which were mainly derived from Zhejiang Province (which called "native species from Zhejiang") and characterized by detection of different degrees of rutin peak, but it was difficult to detect the characteristic peaks of violanthin and isoviolanthin. HPLC characteristic chromatograms of D. officinale in bionics wild cultivation and greenhouse of "Danxia landform species" and "Guangxi Tiepilan species" were compared. The results showed that the characteristic peaks in D. officinale planted in greenhouse could be detected stably,which verified the reliability of the source in D. officinale. Conclusion: The analytical method has a better separation effect on flavonoids of D. officinale, with a good reproducibility. The commonness and specificities of flavonoid glycosides components of D. officinale from different categories have basically confirmed. This suggests that Vicenin Ⅱ is suitable to be a reference peak for characteristic chromatogram. Both the relative abundance of rutin and the detection or relative abundance of violanthin and isoviolanthin peaks could be used as a reference to judge the categories of D. officinale in "Danxia landform species" or "Tiepilan species from Yunnan, South Guangdong and Guangxi" or "native species from Zhejiang".
9. Study of a-Asarone protecting BV2 cells damaged by OGD/R by regulating of NLRP3 pathway
Fei-Fei XU ; Kai GUI ; Li-You WANG ; Ya-Qi GUAN ; Ming LIU ; Qin-Qing LI ; Ya-Juan TIAN ; Wen-Bin HE ; Shi-Feng CHU
Chinese Pharmacological Bulletin 2022;38(8):1209-1218
Aim To evaluate the protective effect of α-asarone on microglials with cerebral ischemia/reperfusion injury by measuring the expression of polar transformation and related inflammatory proteins in BV2 cells in vitro and its mechanisms.Methods The cerebral ischemia/reperfusion injury BV2 cells were pretreated by α-asarone in vitro and simulated by OGD/R model.The effect of α-asarone on the viability of damaged cells in OGD/R model was determined by CCK-8; the morphological changes of cells were observed to analyze the general morphology of cells; the levels of proinflammatory factor IL-1β, IL-18 and anti-inflammatory factor IL-10, IL-4, and ROS activity secreted by BV2 cells were detected by ELISA; the protein expressions of TGF-β, TNF-α and inflammatory related protein NLRP3, caspase 1, p-NF-κB were detected by Western blot.Results The results of in vitro experiments were as follows: the activity of damaged cells in OGD/R model was significantly increased by α-asarone, with the increase of administration dose, the cells in the low, medium and high dose groups of α-asarone decreased, and the "amoeba-like" cells and the cell body were gradually became stereoscopic and full.From the results of cell morphology, it could be seen that α-asarone had a certain proliferative effect on normal cells; the release was significantly reduced of proinflammatory factor IL-1β, IL-18 and TNF-α in OGD/R injured BV2 cells pretreated with α-asarone, also increased the release of IL-10, IL-4 and TGF-β, with a dose-effect relationship, and the high dose(16 μmol·L-1)was the best; the expressions of inflammatory related protein NLRP3, caspase 1, NF-κB and ROS activity in injured cells of OGD/R model were significantly reduced after pretreatment with α-asarone.Conclusions α-asarone has a significant protective effect on cerebral ischemia/reperfusion injury, mainly by regulating ROS activity and inhibiting phosphorylation of NF-κB, in order to reduce the excessive activation of NLRP3 inflammatory corpuscles reducing the secretion of proinflammatory factor IL-1β and IL-18, promoting the secretion of anti-inflammatory factor IL-10 and IL-4, so as to protect cerebral ischemia/reperfusion injury by anti-inflammatory reaction.
10. Memory-improving Substances Basis and Mechanism of Polygalae Radix, Acori Tatarinowii Rhizoma and Its Couplet Medicines
Xiao-qing LI ; Jia-qi ZHAO ; Ya-juan TIAN ; Cheng HAN ; Qin-qing LI ; Shi-feng CHU ; Wen-bin HE
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(3):190-199
Polygalae Radix and Acori Tatarinowii Rhizoma were first recorded in Shennong's Herbal Classic. Both of them can "improve people's memory". Long-term administration can make body light and macrobian. They have often been used as couplet medicines and the core combination of nootropic and memory improvement prescriptions. At present, traditional Chinese medicine clinicians believes that the principle of Polygalae Radix and Acori Tatarinowii Rhizoma in improving memory or intelligence is to supplement the deficiency, remove phlegm and unblock nine orifices, with sufficient evidences for the traditional theory. However, its material basis and mechanism for improving memory have not been fully elucidated. In this paper, we searched the literatures about pharmacological and pharmacodynamics mechanism of Polygalae Radix,Acori Tatarinowii Rhizoma and their chemical components on nervous system in recent ten years from Pubmed database and CNKI. The main material basis for improving memory of Polygalae Radix-saponins, oligosaccharides and alone, the main material basis for improving memory of Acori Tatarinowii Rhizoma-α-asarone,β-asarone and eugenol, the changes of the quality and quantity of the active substances after combination, and the mechanism of improving memory of the single drugs and their couplet medicines, such as scavenging free radicals, regulating cholinergic system, clearing β-amyloid protein(Aβ), decreasing the level of phosphorylation of Tau protein, improving the rate of apoptosis and regulating synaptic plasticity, were systematically collected, analyzed and summarized. In view of the current research situation, this paper points out the possible shortcomings, with the aim to further explore the mechanism of Polygalae Radix combined with Acori Tatarinowii Rhizoma with the mechanism of "1+1>2".