1. Porcine circovirus 3 capsid protein induces autophagy in HEK293T cells by inhibiting phosphorylation of the mammalian target of rapamycin
Journal of Zhejiang University. Science. B 2020;21(7):560-570
Porcine circovirus 3 (PCV3) has been detected in major pig-producing countries around the world since its first report in the US in 2016. Most current studies have focused on epidemiological investigations and detection methods of PCV3 because of lack of live virus strains for research on its pathogenesis in porcine cells or even in pigs. We constructed a recombinant plasmid pCMV-Cap carrying the PCV3 orf2 gene to investigate the effects of capsid (Cap) protein expression on autophagic response in human embryonic kidney cell line 293T (HEK293T). We demonstrate that PCV3 Cap protein induced complete autophagy shown as formation of autophagosomes and autophagosome-like vesicles as well as LC3-II conversion from LC3-I via inhibiting phosphorylation of the mammalian target of rapamycin (mTOR) in HEK293T cells. The ubiquitin-proteasome pathway is also involved in the autophagy process. These findings provide insight for further exploration of PCV3 pathogenetic mechanisms in porcine cells.
3.Erythrocytic or serum hydrogen sulfide association with hypertension development in untreated essential hypertension.
Mei ZHENG ; Qiang ZENG ; Xiao-Qin SHI ; Jing ZHAO ; Chao-Shu TANG ; Ning-Ling SUN ; Bin GENG
Chinese Medical Journal 2011;124(22):3693-3701
BACKGROUNDEndogenous hydrogen sulfide (H(2)S) plays an important role in hypertension. The aim of this study was to investigate the role of erythrocyte and serum H(2)S in patients with untreated essential hypertension.
METHODSWe recruited 62 patients (age 22 - 74 years) with untreated prehypertension or hypertension, and 64 normotensive subjects (age 18 - 64 years). We assessed the 3-mercaptopyruvate sulphurtransferase (MPST) protein expression in erythrocytes and measured the H(2)S production from erythrocytes and serum H(2)S levels, then analyzed the association of erythrocytic or serum H(2)S content and blood pressure or cardiovascular risk factors (e.g., age, body mass index (BMI) and dyslipidemia). A stepwise regression analysis was used to evaluate the possible relationship of erythrocytic H(2)S in hypertension.
RESULTSIn hypertensive patients, erythrocyte H(2)S production ((111.04 ± 29.20) nmol/min per 10(8) erythrocytes) was higher than that in controls ((78.85 ± 19.38) nmol/min per 10(8) erythrocytes), and serum H(2)S was also higher. The erythrocytic H(2)S production was associated with increased systolic blood pressure (sBP), diastolic blood pressure (dBP), age, BMI, level of C-reactive protein (CRP), as well as triglycerides (TG) and high density lipoprotein cholesterol (HDL-C). Serum H(2)S was not associated with age or CRP. Stepwise regression analysis showed that erythrocytic H(2)S production was correlated with sBP, TG, HDL-C, low density lipoprotein cholesterol (LDL-C) and blood urea nitrogen (BUN) and serum H(2)S was correlated with dBP and TG. Results of receiver-operating characteristic curve analysis suggested that erythrocytic H(2)S production was a more sensitive predictor of hypertension development than serum H(2)S.
CONCLUSIONErythrocytic or serum H(2)S production is sensitive predictor of hypertension.
Adolescent ; Adult ; Aged ; Erythrocytes ; metabolism ; Female ; Humans ; Hydrogen Sulfide ; blood ; Hypertension ; blood ; metabolism ; Male ; Middle Aged ; Young Adult
4.Role of sphingosine 1-phosphate receptor signaling in hematopoietic stem/progenitor cell transmigration.
Wen-chao OU ; Shi-ming LIU ; Long-geng XIONG ; Guo-qing LI ; Meng-qun TAN
Journal of Southern Medical University 2009;29(9):1862-1865
OBJECTIVETo determine the role of sphingosine 1-phosphate receptor (S1PRs ) signaling in CD34+ hematopoietic stem/progenitor cell transmigration.
METHODSCD34(+) cells were separated by Ficoll density gradient centrifugation and incubated in DMEM medium with 10% fetal calf serum. The cells were pretreated by FTY720, with or without pertussis toxin (PTX) and antiCXCR4 mAb in the medium, followed by addition of 100 ng/ml SDF-1 into the lower chamber of a Costar 24-well transwell. The migrated cells were counted using FACS and the migrating rates were determined. The expressions of sphingosine 1-phosphate receptors were analyzed in CD34(+) cells before and after the transmigration by reverse transcriptase- polymerase chain reaction (RT-PCR). Cord blood CD34(+) cells were treated with or without FTY720 (10(+) mol/L), and the expressions of CD49d (VLA-4), CD11a (LFA-1), and CD62L (L-selectin) were analyzed at 1, 8, and 16 h after the treatment.
RESULTSWhile FTY720 did not affect spontaneous migration, a substantial increase of SDF-1-induced transmigration was observed in the presence of FTY720 (15.26 2.14 to 28.64 2.37). The FTY720-enhanced transmigration was completely blocked by addition of PTX or antiCXCR4 mAb. S1p1-5 was expressed in fresh isolated cord blood CD34(+) cells. The migrating cells stimulated by FTY720 and SDF-1 only expressed S1P1, S1P3, and S1P4. The expressions of CD49d, CD11a and CD62L on CD34(+) cells treated with FTY720 remained unchanged at the selected time points as compared with the control.
CONCLUSIONSS1PRs are involved the transmigration of CD34(+) cells. The activation of S1PRs results in increased chemotactic response of CD34(+) to SDF-1. These effects are mediated through CXCR4 and PTX-sensitive Gi proteins. Only the CD34(+) cells expressing the specific receptors can rapidly transmigrate. The activation of the S1PRs does not affect the expressions of the adhesion molecules on cord blood CD34(+) cells.
Antigens, CD34 ; metabolism ; Cell Movement ; Cells, Cultured ; Chemokine CXCL12 ; pharmacology ; Fetal Blood ; cytology ; Fingolimod Hydrochloride ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Propylene Glycols ; pharmacology ; Receptors, Lysosphingolipid ; metabolism ; physiology ; Signal Transduction ; Sphingosine ; analogs & derivatives ; pharmacology
5.Clinical evaluation of wedge-shaped defect restored by gingival retraction technique.
Wei-jiang YANG ; Geng-sheng SHI ; Li-rong LIANG ; Fang-chao RUAN ; Hui-juan GAO ; Yong-qing TONG
Chinese Journal of Stomatology 2007;42(6):353-354
OBJECTIVETo evaluate the periodontal conditions after the wedge-shaped defect was restored by gingival retraction technique.
METHODSA total of 138 mandibular premolars with wedge-shaped defect were selected and divided into A, B groups. Group A was restored with Dyract after using retraction cord. Group B was directly restored with Dyract. Clinical parameters including plaque index (PLI), gingival index (GI), sulcus bleeding index (SBI), probing depth (PD), volumes of gingival crevicular fluid (GCF) and levels of aspartate aminotransferases (AST) of gingival crevicular fluid were measured at baseline, 1 week, 1 month, 3 months and 6 months after operation.
RESULTSThere was no difference in PLI, GI, SBI, PD between group A and B during 6 months after operation, while the difference of GCF and AST was significant between group A and B at 3 months and 6 months after operation (P < 0.05, P < 0.01).
CONCLUSIONSGingival retraction technique applied in wedge-shaped defect restoration can reduce the damage to the periodontal tissue.
Adolescent ; Adult ; Aspartate Aminotransferases ; analysis ; Dental Plaque Index ; Dental Restoration, Permanent ; methods ; Female ; Gingival Crevicular Fluid ; enzymology ; Humans ; Male ; Periodontal Index ; Young Adult
6.Porcine circovirus 3 capsid protein induces autophagy in HEK293T cells by inhibiting phosphorylation of the mammalian target of rapamycin.
Shi-Chao GENG ; Xiao-Liang LI ; Wei-Huan FANG
Journal of Zhejiang University. Science. B 2020;21(7):560-570
Porcine circovirus 3 (PCV3) has been detected in major pig-producing countries around the world since its first report in the US in 2016. Most current studies have focused on epidemiological investigations and detection methods of PCV3 because of lack of live virus strains for research on its pathogenesis in porcine cells or even in pigs. We constructed a recombinant plasmid pCMV-Cap carrying the PCV3 orf2 gene to investigate the effects of capsid (Cap) protein expression on autophagic response in human embryonic kidney cell line 293T (HEK293T). We demonstrate that PCV3 Cap protein induced complete autophagy shown as formation of autophagosomes and autophagosome-like vesicles as well as LC3-II conversion from LC3-I via inhibiting phosphorylation of the mammalian target of rapamycin (mTOR) in HEK293T cells. The ubiquitin-proteasome pathway is also involved in the autophagy process. These findings provide insight for further exploration of PCV3 pathogenetic mechanisms in porcine cells.
7.Study on the epidemiological characteristics and natural infectious focus of Angiostrongylus cantonensis in Shenzhen area of Zhujiang Delta in China
Ren-Li ZHANG ; Shi-Tong GAO ; Yi-Jie GENG ; Da-Na HUANG ; Mu-Xin CHEN ; Jian-Ping LIU ; Yuan-Liang WU ; Yin ZHEN ; Zhuan-Wen DAI ; Qi-Wen ZHANG ; Tai-Shun WU ; Zhi-Chao MA ; Wu-Sheng CHEN ; Da-Lin LI
Chinese Journal of Epidemiology 2008;29(6):573-576
Objective To delimit the natural infectious focus, including the distribution of wildlife,species, ecology of intermediate hosts and final host of Angiostrongylus cantonensis, as well as the routes of transmission and epidemiological characteristics and wildlife of human Angiostrongylus cantonensis, based on human diverging cases identified in Shenzhen, southern area of China. Methods Data including rate of infection and density of Angiostrongylus cantonensis among different hosts in 12 different areas in Shenzhen was collected, using microscope to inspect homogenate liquids of snails. Wild mice were captured with mouse cage to examine the adult Angiostrongylus cantonensis. Using larva isolated from wild-snails-infected rats to observe the life cycle of Angiostrongylus cantonensis. Results Wild life of Angiostrongylus cantonensis existed in the southwest part of Shenzhen with its majority intermediate hosts as Achatina fulica. The overall rate of infection was 31% in wildlife and final host was found to be Rattus andersoni, Achatina fulica which were extensively distributed in the shrub region of Shenzhen because of suitable climate,humidity and vegetation for generating the life cycle of Achatina fulica. Human infected Angiostrongylus cantonensis was mainly due to eating raw snails or vegetables contaminated by larva of Angiostrongylus cantonensis.The peak of infection was seen from April to November in Shenzhen area.Conclusion Wildlife of Angiostrongylus cantonensis existed in the southwest part of Shenzhen with major wildlife reservoir including fresh water snail and wild mouse. The existence of natural focus Angiostrongylus cantonensis was now recognized as an important source of human angiostrongliasis in Shenzhen area.
8.The Serum Exosome Derived MicroRNA-135a, -193b, and -384 Were Potential Alzheimer's Disease Biomarkers.
Ting Ting YANG ; Chen Geng LIU ; Shi Chao GAO ; Yi ZHANG ; Pei Chang WANG
Biomedical and Environmental Sciences 2018;31(2):87-96
OBJECTIVE:
MicroRNAs (miRs) are attractive molecules to be considered as one of the blood-based biomarkers for neurodegenerative disorders such as Alzheimer's disease (AD). The goal of this study was to explore their potential value as biomarkers for the diagnosis of AD.
METHODS:
The expression levels of exosomal miR-135a, -193b, and -384 in the serum from mild cognitive impairment (MCI), dementia of Alzheimer-type (DAT), Parkinson's disease with dementia (PDD), and vascular dementia (VaD) patients were measured with a real-time quantitative reverse transcriptase PCR (qRT-PCR) method.
RESULTS:
Both serum exosome miR-135a and miR-384 were up-regulated while miR-193b was down-regulated in serum of AD patients compared with that of normal controls. Exosome miR-384 was the best among the three miRs to discriminate AD, VaD, and PDD. Using the cut-off value could better interpret these laboratory test results than reference intervals in the AD diagnosis. ROC curve showed that the combination of miR-135a, -193b, and -384 was proved to be better than a particular one for early AD diagnosis.
CONCLUSION
Our results indicated that the exosomal miRs in the serum were not only potential biomarker of AD early diagnosis, but might also provide novel insights into the screen and prevention of the disease.
Aged
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Alzheimer Disease
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blood
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Biomarkers
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blood
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Case-Control Studies
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Cognitive Dysfunction
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blood
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Dementia, Vascular
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blood
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Diagnosis, Differential
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Early Diagnosis
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Exosomes
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metabolism
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Female
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Humans
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Male
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MicroRNAs
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blood
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Parkinson Disease
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blood
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Sensitivity and Specificity
9.Effects of Tc17 Cells on Disease Severity and HBV-DNA Elimination in Patients with HBV Related Acute-on-chronic Liver Failure
Geng-lin ZHANG ; Shi-cheng XU ; Ting ZHANG ; Chao-shuang LIN ; Liang PENG ; Zhi-liang GAO
Journal of Sun Yat-sen University(Medical Sciences) 2019;40(5):739-746
【Objective】To investigate the effects of Tc17 cells on disease severity and elimination of HBV-DNA in
patients with HBV related acute-on-chronic liver failure(HBV-ACLF).【Methods】Forty-three patients with HBV-
ACLF were enrolled. Twenty patients with chronic hepatitis B(CHB)and 12 healthy subjects(NC)were enrolled as
controls. Flow cytometry was used to detect the expression of peripheral Tc17 cells. HBV-DNA loads were measured,and
the MELD,MELD-Na,CLIF-C ACLF and AARC scores were used to evaluate the disease severity. The effects of Tc17
cells on disease severity and decline of HBV-DNA were analyzed.【Results】Compared with NC group and CHB group,
the expression of Tc17 cells in HBV-ACLF patients was significantly increased(P < 0.001 and P = 0.017). Correlation
analysis showed that Tc17 cells were positively correlated with AARC score,MELD score and MELD-Na score(r =
0.504,P = 0.001;r = 0.417,P = 0.005 and r = 0.382,P = 0.012),and a correlated trend was found with CLIF-C
ACLF score(r = 0.294,P = 0.055). And as the disease progressed,the expression of Tc17 cells gradually increased.In addition,Tc17 cells were positively correlated with HBV-DNA levels(r = 0.339,P = 0.026)at baseline,and the
HBV-DNA levels were significantly decreased in patients with higher expression of Tc17 cells than in lower group after 4
weeks of treatment(P < 0.001). Furthermore,it was found that baseline Tc17 cells and AST levels were associated with
the HBV-DNA decline by multivariate linear regression analysis.【Conclusion】Tc17 cells-mediated inflammatory
response helps clear the HBV-DNA,but excessive inflammatory response may aggravate the disease severity.
10.Expert consensus on the diagnosis and treatment of osteoporotic proximal humeral fracture with integrated traditional Chinese and Western medicine (version 2024)
Xiao CHEN ; Hao ZHANG ; Man WANG ; Guangchao WANG ; Jin CUI ; Wencai ZHANG ; Fengjin ZHOU ; Qiang YANG ; Guohui LIU ; Zhongmin SHI ; Lili YANG ; Zhiwei WANG ; Guixin SUN ; Biao CHENG ; Ming CAI ; Haodong LIN ; Hongxing SHEN ; Hao SHEN ; Yunfei ZHANG ; Fuxin WEI ; Feng NIU ; Chao FANG ; Huiwen CHEN ; Shaojun SONG ; Yong WANG ; Jun LIN ; Yuhai MA ; Wei CHEN ; Nan CHEN ; Zhiyong HOU ; Xin WANG ; Aiyuan WANG ; Zhen GENG ; Kainan LI ; Dongliang WANG ; Fanfu FANG ; Jiacan SU
Chinese Journal of Trauma 2024;40(3):193-205
Osteoporotic proximal humeral fracture (OPHF) is one of the common osteoporotic fractures in the aged, with an incidence only lower than vertebral compression fracture, hip fracture, and distal radius fracture. OPHF, secondary to osteoporosis and characterized by poor bone quality, comminuted fracture pattern, slow healing, and severely impaired shoulder joint function, poses a big challenge to the current clinical diagnosis and treatment. In the field of diagnosis, treatment, and rehabilitation of OPHF, traditional Chinese and Western medicine have accumulated rich experience and evidence from evidence-based medicine and achieved favorable outcomes. However, there is still a lack of guidance from a relevant consensus as to how to integrate the advantages of the two medical systems and achieve the integrated diagnosis and treatment. To promote the diagnosis and treatment of OPHF with integrated traditional Chinese and Western medicine, relevant experts from Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine have been organized to formulate Expert consensus on the diagnosis and treatment of osteoporotic proximal humeral fracture with integrated traditional Chinese and Western medicine ( version 2024) by searching related literatures and based on the evidences from evidence-based medicine. This consensus consists of 13 recommendations about the diagnosis, treatment and rehabilitation of OPHF with integrated traditional Chinese medicine and Western medicine, aimed at standardizing, systematizing, and personalizing the diagnosis and treatment of OPHF with integrated traditional Chinse and Western medicine to improve the patients ′ function.