1. Effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress
Ben-Wen ZHOU ; Chang-Cheng ZHANG ; He DENG ; Si-Min CHEN ; Yan-Yu CHANG ; Yan-Na YANG ; Guo-Qing FU ; Ding YUAN ; Hai-Xia ZHAO ; Ben-Wen ZHOU ; He DENG ; Yan-Na YANG ; Hai-Xia ZHAO ; Ben-Wen ZHOU ; Chang-Cheng ZHANG ; Si-Min CHEN ; Yan-Yu CHANG ; Guo-Qing FU
Chinese Pharmacological Bulletin 2023;39(12):2346-2353
Aim To study the effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress. Methods C57BL/6J male mice were assigned into normal group and high-fat diet group randomly, with six mice in each group. The mice in normal group or high-fat diet group were fed with regular or high-fat diet continuously for five months. The mice were weighed, anesthetized, and euthanized to collect testicular and epididymal tissue for analysis. The testicular tissue was weighed and their indices were calculated. Epididymal tissue was collected for semen analysis. The morphological alterations of testicular tissue were observed using hematoxylin-eosin ( HE ) staining. The apoptosis of germ cells was detected by TUNEL staining and the apoptotic indices were calculated. The expression levels of apoptosis and endoplasmic reticulum stress-related proteins in testicular tissue were detected by Western blot. The protein expression and localization of GRP78 in testicular tissue were further detected by immunofluorescence. Results The results showed that compared to the normal group, the high-fat diet group had a significant increase in body weight, a significant decrease in testicular index, sperm concentration, and sperm vability, loose arrangement of germ cells, significant thinning of the seminiferous epithelium, no significant change in the diameter of seminiferous tubules, a significant increase in germ cell apoptosis , with an increased apoptosis index, and significant increase in expression of Bax and cleaved-caspase-12,and a significant decrease in Bcl-2 protein expression. The expression levels of GRP78 , p-IREl, XBP1, and ATF6a proteins were significantly up-regulated, while p-PERK, p-eIF2a, ATF4 protein expression showed no significant changes. Immunofluorescence results further showed a significant increase in the expression of GRP78 protein in the testicular tissue,with no significant changes in the expression location. Conclusions High-fat diet can induce the apoptosis of mouse testicular germ cells, and the mechanism may be related to the activation of endoplasmic reticulum stress IRE1 and ATF6 signaling pathway.
2.Huangqi Decoction, a compound Chinese herbal medicine, inhibits the proliferation and activation of hepatic stellate cells by regulating the long noncoding RNA-C18orf26-1/microRNA-663a/transforming growth factor-β axis.
Ben-Sheng DONG ; Fu-Qun LIU ; Wen-Na YANG ; Xiao-Dong LI ; Miao-Juan SHI ; Mao-Rong LI ; Xiu-Li YAN ; Hui ZHANG
Journal of Integrative Medicine 2023;21(1):47-61
OBJECTIVE:
Huangqi Decoction (HQD), a classical traditional Chinese medicine formula, has been used as a valid treatment for alleviating liver fibrosis; however, the underlying molecular mechanism is still unknown. Although our previous studies showed that microRNA-663a (miR-663a) suppresses the proliferation and activation of hepatic stellate cells (HSCs) and the transforming growth factor-β/small mothers against decapentaplegic (TGF-β/Smad) pathway, whether long noncoding RNAs (lncRNAs) are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported. The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.
METHODS:
The expression levels of lnc-C18orf26-1, miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction. HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs. The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs. Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs. RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs. Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA. The expression levels of collagen α-2(I) chain (COL1A2), α-smooth muscle actin (α-SMA) and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.
RESULTS:
Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a. Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation, downregulated TGF-β1-stimulated α-SMA and COL1A2 expression, and inhibited the TGF-β1/Smad signaling pathway. HQD suppressed the proliferation and activation of HSCs. HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs. Further studies showed that HQD inhibited the expression of COL1A2, α-SMA, TGF-β1, TGF-β type I receptor (TGF-βRI) and phosphorylated Smad2 (p-Smad2) in HSCs, and these effects were reversed by miR-663a inhibitor treatment.
CONCLUSION
Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis. HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.
Humans
;
Transforming Growth Factor beta/pharmacology*
;
Transforming Growth Factor beta1/metabolism*
;
RNA, Long Noncoding/pharmacology*
;
Drugs, Chinese Herbal/pharmacology*
;
MicroRNAs/genetics*
;
Hepatic Stellate Cells/pathology*
;
Liver Cirrhosis/metabolism*
;
Cell Proliferation
;
Transforming Growth Factors/pharmacology*
3.Prevalence of human soil-borne nematode infections in Yunnan Province: a cross-sectional study in 2015
Fang-Wei WU ; Li-Bo WANG ; Ben-Fu LI ; Xin-Liu YAN ; Jin-Rong ZI ; Jia PENG ; Xuan CAI ; Xue-Ying BAO ; Ya-Ming YANG
Chinese Journal of Schistosomiasis Control 2021;33(5):513-517
Objective To investigate the current prevalence of human soil-borne nematode infections in Yunnan province, so as to provide the scientific evidence for formulating the soil-borne nematodiasis control strategy in the province. Methods In 2015, a total of 20 survey sites were sampled in 10 counties (cities) of Yunnan Province using the stratified cluster random sampling method. Stool samples were collected from all local permanent residents at ages of one year and older in each survey site, and the soil-borne nematode eggs were identified using the modified Kato-Katz technique and the egg number was counted. In addition, the hookworm species was identified using the filter-paperculture method, and Enterobius vermicularis eggs were detected using the cellophane tape method in children at ages of 3 to 6 years. Results A total of 5 067 residents received stool examinations, and 950 residents were detected with soil-borne nematode infections, with an overall prevalence rate of 18.75%. The prevalence of Ascaris lumbricoides, Trichuris trichura and hookworm was 7.52%, 8.47% and 9.02%, respectively. Among 446 children detected using the cellophane tape method, 5 children were detected with E. vermicularis infections. Among the 160 residents with hookworm infections, there were 139 residents with Necator americanus infections (86.88%), 16 with A. duodenale infections (10.00%) and 5 with mixed infections (3.12%). Mild A. lumbricoides (67.98%, 259/381), T. trichura (88.58%, 380/429) and hookworm infections (94.53%, 432/457) were predominant. Among the four ecological zones, the highest prevalence of human soilborne nematode infections was found in the East Tibet-South Sichuan Ecological Zone (31.79%), and among the 10 survey counties (cities), the greatest prevalence was seen in Gongshan Derung and Nu Autonomous County (50.13%), while the lowest prevalence was found in Ninglang Yi Autonomous County (0.40%). The prevalence of human soil-borne nematode infections was 5.67% (43/759), 26.67% (610/2 287) and 14.70% (297/2 021) in high-, moderate- and low-economic-level regions, respectively. There were no significant differences in the prevalence of human soil-borne nematode infections in terms of ecological regions, survey counties (cities) or economic development levels (χ2 = 342.20, 814.60 and 201.34, all P < 0.05). There was no significantdifference in the prevalence of human soil-borne nematode infections between male (18.21%, 441/2 422) and female residents (19.24%, 509/2 645) (χ2 = 0.89, P > 0.05), and soil-borne nematode infections were detected in residents at all age groups, with the greatest prevalence found in residents at ages of 1 to 9 years (25.88%). In addition, the highest prevalence of soil-borne nematode infections was seen in residents with the Dulong Ethnic Minority (82.09%), in preschool children (25.06%) and in illiterate residents (24.80%), and there was no age-, ethnicity-, occupation- or education level-specific prevalence of soil-borne nematode infections detected (χ2 = 46.50, 1 016.96, 36.33 and 52.43, all P < 0.05). Conclusions The prevalence of human soil-borne nematode infections remains high in Yunnan Province. The management of soil-borne nematodiasis requires to be reinforced among low-age children, farmers, old people and residents with low educations levels or ethnic groups.
4.Allelopathic Effect of Water Extracts from Rhizosphere Soil of Three Medicinal Plants on Polygala tenuifolia Seeds and Their Seedlings
Rui-xiang LEI ; Bing-yue YANG ; Yan-ni WEI ; Fu-lin CAO ; Yi-cong FAN ; Sha-sha WU ; Gang ZHANG ; Yong-gang YAN ; Liang PENG ; Ben-xiang HU
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(23):161-170
Objective:To investigate the allelopathic effects of water extracts from rhizosphere soil of three medicinal plants
5.Analysis on Polymorphism of Platelet Antigen Gene in Shandong Han Population.
Yan LIU ; Hai-Feng ZHU ; Wen-Ben QIAO ; Yi ZHANG ; Yong-Hong SONG ; Xiang-Min NIE ; Chuan-Fu ZHU ; Pei-Cong ZHAI
Journal of Experimental Hematology 2020;28(6):2066-2070
OBJECTIVE:
To study the Polymorphism of the human platelet antigen(HPA) gene 1-17 and human leukocyte antigen(HLA) gene-A and B locus in Shandong Han population.
METHODS:
A total of 962 samples from routine voluntary platelet donors were genotyped for HPA1-17 system and HLA-A site, B by PCR-SSP and PCR-SSOP respectively.Gene frequencies were calculated by counting. HPA1-17 and HLA genotype combinations were analyzed by Arelequin 3.5.
RESULTS:
The gene frequencies of HPA-la, -1b, HPA-2a, -2b, HPA-3a, -3b, HPA-4a, -4b, HPA-5a, -5b, HPA-6a, -6b, HPA-15a, -15b were 0.9918, 0.0082, 0.9419, 0.0592, 0.5841, 0.4174, 0.9969, 0.0031, 0.9892, 0.0108, 0.9835, 0.0175,0.5488 and 0.4512, respectively. The most common HPA genotype combination was HPA-(1, 2, 4, 5, 6, 7-14, 16, 17) aa-3ab-15ab (0.2048). Moreover, HLA-A*2(0.3094) and HLA-B*13(0.1513) showed the highest frequency in their respective locus. The most common HLA genotype combination was HLA-A*2-B*13(0.1397) .
CONCLUSION
Distributions of HPA and HLA show high polymorphism in Shandong Han population. The ethnic and territorial difference of HPA distribution is also confirmed. It is imperative to establish local genetic database of volunteer platelet donors.
Alleles
;
Antigens, Human Platelet/genetics*
;
Gene Frequency
;
Genotype
;
Humans
;
Polymorphism, Genetic
6. Effect of Hydrogen Peroxide (H2O2) on Content of Total Phenol Flavonoids and Activity of Related Enzymes in Polygala tenuifolia Callus
Fu-lin CAO ; Ben-xiang HU ; Liang PENG ; Lu LUO ; Jie LI ; Ting ZHAO ; Yan-ru AN ; Tao HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(20):153-159
Objective:To study the changes in total phenolic and flavonoid content and antioxidant enzyme activity of Polygala tenuifolia callus in MS medium with different concentrations of H2O2,in order to explore the physiological mechanism of Polygala tenuifolia callus in adapting to H2O2 environmental stress at the cellular level. Method:Five gradients of 0,5,10,15,20 mmol·L-1 were set for H2O2 concentration and added to MS medium, with P. tenuifolia callus as the experimental material. Total phenols,total flavonoids and antioxidant enzyme activities of callus were determined after 5,10,15,20,25 d of culture,respectively. Result:The contents of total phenols and flavonoids were the highest when the concentration of H2O2 was 5 mmol·L-1 for 15 d. The SOD activity was the highest when the callus was cultured for 5 d and the exogenous H2O2 concentration was 5 mmol·L-1. POD activity was the highest at 25 d and 5 mmol·L-1 concentration of exogenous H2O2.CAT activity was the highest at 25 d and 15 mmol·L-1 concentration of exogenous H2O2. Conclusion:P. tenuifolia callus has the ability to adapt to the environmental stress of H2O2 at a certain concentration. When it is subjected to the environmental stress of H2O2,P. tenuifolia callus can alleviate the damage by regulating its secondary metabolites and protecting enzyme system. It can significantly promote the content of total phenols and flavonoids in secondary metabolites at 5-10 mmol·L-1. SOD activity was significantly increased at 5 d and the concentration of exogenous H2O2 of 5 mmol·L-1. POD activity was significantly increased at 25 d and the concentration of exogenous H2O2 of 5 mmol·L-1. CAT activity was significantly increased at 25 d and concentration of exogenous H2O2 of 15 mmol·L-1.
7. Effect of Different Hormone Ratios on Callus Induction and Flavone Accumulation in Polygala tenuifolia
Jie LI ; Ben-xiang HU ; Liang PENG ; Bing-yue YANG ; Lu LUO ; Fu-lin CAO ; Ting ZHAO ; Yan-ru AN ; Tao HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(19):131-137
Objective:To study the effect of different hormone ratios on the callus induction of roots,stems and leaves of Polygala tenuifolia,and determine and analyze the amount of flavonoids in roots,stems and leaves of P. tenuifolia. Method:With MS as the basic medium and roots,stems and leaves of P. tenuifolia sterile seedings as explants,the effects of 2,4-D,NAA and 6-BA on callus induction and flavonoid accumulation in different parts of roots,stems and leaves of P. tenuifolia were determined by orthogonal test. Result:2,4-D,NAA and 6-BA had significant effects on the callus induction rate of roots,stems and leaves of P. tenuifolia. The optimal callus induction combination of leaves was MS+3.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.5 mg·L-1 6-BA,the optimal callus induction combination of stems was MS+1.0 mg·L-1 2,4-D+3.0 mg·L-1 NAA+1.5 mg·L-1 6-BA,the optimal callus induction combination for roots was MS+1.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.0 mg·L-1 6-BA. And 2,4-D,NAA and 6-BA had significant effects on flavonoid accumulation in the stem callus of P. tenuifolia,and MS+3.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+0.5 mg·L-1 6-BA was the best flavonoid accumulation combination.NAA,6-BA had significant effects on flavonoid accumulation in the leave callus of P. tenuifolia,while 2,4-D had no significant effect,and MS+3.0 mg·L-1 2,4-D+2.0 mg·L-1 NAA+1.0 mg·L-1 6-BA was the optimal flavonoid accumulation combination,the three hormones had no significant effect on the accumulation of flavonoids in the root callus of P. tenuifolia,and MS+2.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.5 mg·L-1 6-BA was the best flavonoid accumulation combination. Conclusion:Under the conditions,the callus induction rate of roots,stems and leaves of P. tenuifolia is 100%, especially, the callus of P. tenuifolia leaves was the optimal,which is followed by P. tenuifolia stems and P. tenuifolia roots. Under the conditions,the amount of flavonoids in roots,stems and leaves of P. tenuifolia reach 21.31,24.56,23.61 mg·g-1,respectively.
8.Analysis of influencing factors of secondary metabolites contents in cultivated Polygala tenuifolia.
Ya-Jie PU ; Dan-Dan WANG ; Yan YAN ; Hong-Ling TIAN ; Bing PENG ; Xue-Mei QIN ; Cun-Gen MA ; Chen-Hui DU ; Ben-Xiang HU ; Fu-Sheng ZHANG
China Journal of Chinese Materia Medica 2017;42(16):3167-3177
This work was launched to explore the effect of habitat and growth year on the secondary metabolites contents of cultivated Polygala tenuifolia. The samples of cultivated P. tenuifolia were analyzed by ultra-high performance liquid chromatography(UPLC)-quadrupole time-of-flight mass spectrometry(Q-TOF MS), and the obtained data were analyzed using multiple statistical analysis and cluster analysis. The results showed that compared with growth year, habitat is a main influencing factor which affected the secondary metabolites contents of P. tenuifolia. The contents of sucrose esters and oligosacchride multi-esters are greatly dependent on the habitat (the sample-AG with high levels of components of tenuifoliside B and tenuifoliside C, and the sample-FY with high levels of 3,6'-disinapoyl sucrose, tenuifoliose S, tenuifoliose L, and tenuifoliose V). There is no obvious effect of habitat and growth year on xanthone. The contents of triterpene saponins are greatly dependent on the growth year, and the content of parts of triterpene saponins increased as time goes on.The result indicated that the effect of habitat and growth year on different types of secondary metabolites is not completely equivalent. This study will contribute to the breeding of P. tenuifolia and amendment of current commodity criteria.
9.Identification of a novel HLA allele, HLA-DRB1*03:80, by sequencing-based typing.
Xiang-Min NIE ; Yi ZHANG ; Yun-Long ZHUANG ; Yong-Hong SONG ; Wen-Ben QIAO ; Yan LIU ; Chuan-Fu ZHU
Journal of Experimental Hematology 2014;22(2):509-512
This study was aimed to identify a novel HLA-DRB1 allele from a Chinese potential hemopoietic stem cell donor of Northeast China. A rare HLA-DRB1 allele was initially detected by Luminex PCR-SSO typing, then the sample was sequenced by sequence-based typing (SBT) and the alignments of sample's alleles was identified by single allele-specific sequencing strategy. The results revealed the existence of a new allele which differs from the closest matching allele DRB1*03:06 by a single nucleotide substitution at position 239, where C→G in exon 2, resulting in an amino acid exchange from Thr to Arg at codon 51. It is concluded that a novel allele has been confirmed and its name DRB1*03:80 is officially assigned by the WHO Nomenclature Committee in February 2012.
Alleles
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Asian Continental Ancestry Group
;
genetics
;
HLA-DRB1 Chains
;
genetics
;
Humans
;
Male
;
Sequence Analysis, DNA
10.Gefitinib enhances the radiosensitivity of nasopharyngeal carcinoma cell line CNE2 in vitro.
Ben-fu HE ; Ai-min SUN ; Bi-yan HUANG ; Wen-jun WANG ; Xiao-kang ZHENG ; Rong-cheng LUO
Journal of Southern Medical University 2011;31(6):991-994
OBJECTIVETo study the radiosensitizing effect of gefitinib on nasopharyngeal carcinoma cell line CNE2 in vitro.
METHODSNasopharyngeal carcinoma cell line CNE2 was cultured in RP2MI 1640. MTT assay was performed to evaluate the cell proliferation changes in response to gefitinib treatment and the radiosensitizing effect of gefitinib. The cell survival curves and sensitive enhancement ratio (SERs) were obtained with a clonogenic assay. Flow cytometry analysis was applied to detect the cell cycle changes and cell apoptosis.
RESULTSMTT assay showed that cells exposed to gefitinib and radiation had a significantly lower survival ratio compared to the cells with radiation exposure only (0.582∓0.012 vs 0.398∓0.016, P=0.002), with a SER of 1.535∓0.134. The S phase cell percentage was significantly decreased and G(2)-M phase cells increased in gefitinib plus radiation group (P=0.000), suggesting a synergistic effect of gefitinib and radiation.
CONCLUSIONGefitinib can enhance the radiosensitivity of nasopharyngeal carcinoma CNE2 cells in vitro possibly by inhibiting cell proliferation, inducing cell apoptosis, and causing changes in the cell cycle distribution.
Apoptosis ; drug effects ; Carcinoma ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Humans ; Nasopharyngeal Neoplasms ; pathology ; Quinazolines ; pharmacology ; Radiation Tolerance ; drug effects

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