1.Inhibitory effect of lead acetate on TRPA1 channel in mice and humans
Chinese Journal of Pharmacology and Toxicology 2016;30(9):949-954
OBJECTIVE To investigate the inhibitory effect of lead acetate on transient receptor potential A1(TRPA1)channel. METHODS TRPA1-mediated calcium influx in mice dorsal root ganglion(DRG) neurons and HEK293 cells expressing nouse TRP1 (mTRPA1) and human TRPA1 (hTRPA1) was recorded by intracellular calcium imaging. TRPA1-mediated currents were detected by two-electrode voltage clamp. RESULTS Lead acetate 3.0 and 10.0μmol·L-1 inhibited external calcium influx in DRG neurons by(36.7 ± 4.1)% and(79.4 ± 3.1)%(n=5),respectively. The inhibitory effect of lead acetate on hTRPA1-mediated current was concentration-dependent. Lead acetate 0.3, 1.0, 3.0, 10.0 and 30.0μmol · L-1 inhibited the amplitudes of currents by(1.0 ± 0.7)%,(11.6 ± 0.8)%,(57.7 ± 3.2)%,(93.6 ± 2.6)%and(91.2±2.0)%(n≥4),respectively,with the IC50 2.4μmol·L-1. CONCLUSION TRPA1 channel may be an endogenous target of lead. Lead acetate inhibits TRPA1 channel at a very low concentration.
2.Dexmedetomidine alleviates lung ischemia-reperfusion injury through CHOP pathway in mice
Dan CHEN ; Dong SONG ; Yuzhu YE ; Jinbo HE ; Lei CHEN ; Xiaoxiao QIU ; Lina LIN ; Wantie WANG
Chinese Journal of Pathophysiology 2015;(6):1093-1098
[ ABSTRACT] AIM:To explore the effect of dexmedetomidine ( DEX) on the CCAAT/enhancer-binding protein-homologous protein ( CHOP) pathway during lung ischemia-reperfusion ( I/R) in mice.METHODS:C57BL/6J male mice were randomly divided into sham operation group ( sham group) , lung ischemia/reperfusion group ( I/R group) , ischemia/reperfusion +normal saline group ( I/R+NS group ) and ischemia/reperfusion+dexmedetomidine group ( I/R+DEX group) .Dexmedetomidine was infused intraperitoneally with 25 μg/kg for 30 min prior to the ischemia period in I/R+DEX group, the normal saline was administrated with the same volume of dexmedetomidine in I/R+NS group.After fini-shed the 3 h-reperfusion period , the left lung tissues were harvested to determine lung wet/dry weight ( W/D) , the total lung water content ( TLW) , and index of quantitative evaluation for alveolar damage ( IQA) .Morphological observation and terminal-deoxynucleotidyl transferase mediated nick end labeling ( TUNEL) were applied to evaluate the structure changes and the apoptosis index (AI) of the lung tissues.The expression of CHOP and glucose-regulated protein 78 (GRP78) at mRNA and protein levels in the lung tissues was detected by Western blot and RT-PCR.RESULTS:Compared with sham group, the W/D, TLW, IQA, AI, the mRNA and protein expression of CHOP and GRP78 obviously increased, and the left lung tissues structure were damaged more obviously both in I/R group and I/R+NS group.Compared with I/R group, the W/D, TLW, IQA, AI and the protein and mRNA expression of CHOP in I/R+DEX group decreased, the injury of the left lung tissue structures induced by I/R in I/R+DEX group were also alleviated .CONCLUSION:DEX alleviates the
lung I/R injury, which may be related to inhibition of apoptosis mediated by CHOP pathway.
3.CT features of pulmonary mucormycosis in post-hematopoietic stem cell transplantation patients with leukemia
Yuzhu SHI ; Pengfei WANG ; Jie SONG ; Wenqian YU ; Chunli WANG ; Yanmin LUO
Journal of Practical Radiology 2017;33(6):551-553,580
Objective To summarize the CT features of pulmonary mucormycosis in post hematopoietic stem cell transplantation (HSCT) patients with leukemia,to provide timely and accurate guidance for clinical treatment.Methods 9 pulmonary mucormycosis in post HSCT patients with leukemia confirmed by surgery,biopsy and sputum culture were analyzed retrospectively.Distribution,morphology and CT features of the disease were analyzed and summarized.All patients were underwent non-enhanced MSCT.Results Reversed halo sign (n=7);patchy ground glass opacity (GGO) (n =5);bilateral multiple pulmonary nodules and nodular GGO (n=3);bilateral multiple pulmonary micro-cysts with spiculate boundary (n=1);pleural effusion (n=2);pneumo-mediastinum (n 1) were seen.Two or more than two CT signs were seen in some patients.The interval time between the appeared reversed halo sign and the transplantation were 0.5-19 months in 7 patients,which median time was 10 months and 6 cases (85.7%) appeared within 18months.8 patients (88.9 %) presented cough,expectorated white or yellow phlegm,among them,4 cases (50 %) presented bloodstained sputum or hemoptysis.3 cases (37.5 %) presented left chest pain.1 case was asymptomatic.6 patients (66.7 %) presented significant increased body temperature with the range from 38.4 ℃ to 39.6 C,the median was 39 C.3 patients (33.3%) showed normal temperature.The WBC counting was between 0.16-3.31 × 109/L,the median was 0.7 × 109/L.The neutrophil cell counting was between 0-2.46 × 109/L,with the median of 0.09 × 109/L.Among them,7 patients (77.8%) found reversed halo sign with neutrophil counting between 0-1.63 × 109/L,and the median was 0.09 × 109/L.Conclusion Various imaging manifestations of pulmonary mucormycosis are seen in post HSCT patients with leukemia.When patients is in agranulocytosis phase after transplantation HSCT,it is highly suggested pulmonary mucormycosis infection when the characteristic reversed halo sign is found.
4.Establishment and identification of HUVEC cell strains with over-expression and low expression of RACK 1
Li ZHANG ; Xiongfei JIA ; Hua NIU ; Yue FENG ; Yuzhu SONG ; Wanglong ZHANG ; Shengying LI ; Mingjiang WU ; Xiaoqin MAO
Basic & Clinical Medicine 2015;(2):218-223
Objective To establish several human umbilical vein endothelial cell ( HUVEC ) strains with over-ex-pression or low expression of receptor for activated C kinase 1 ( RACK1 ) , which will provide an effective tool for future studying the function of RACK1 in arrhythmia.Methods The full-length cDNA sequence of RACK1 gene was amplified and inserted into pIRES2-EGFP.At the same time, designed and synthesised complementary DNA sequences of 3 pairs of short hairpin structure and a pair of negative control sequence , then subcloned into the plas-mid pGenesil-1 .The HUVEC cells were transfected with these plasmids and screened by using G 418 .And the expression of RACK1 mRNA and protein in the cells were assayed by qRT-PCR and Western blot , respectively . Results RACK1 eukaryotic expression vector and siRNA expression vectors of RACK 1 were constructed success-fully.After a 48 h transfection of HUVEC cells with the recombinant vectors and G 418 selection, the positive cell clones were obtained .qRT-PCR and Western blot showed that over-expression vector and interference vectors could effectively enhanced and knocked-down RACK1 expression in HUVEC strains .Conclusions HUVEC cell strains with over-expression and low expression of RACK 1 have been successfully established .
5.MRI manifestations of immunosuppressive drugs associated encephalopathy after allogeneic hematopoietic stem cell transplantation in children with aplastic anemia
Yuzhu SHI ; Xinyu YUAN ; Yuchun YAN ; Jie SONG ; Chunli WANG ; Yanmin LUO ; Jiaman WEI
Chinese Journal of Radiology 2017;51(10):782-785
Objective To summarize the MRI manifestations of immunosuppressive drugs associated encephalopathy after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with aplastic anemia. Methods Fifteen patients with immunosuppressive drugs associated encephalopathy after allogeneic hematopoietic stem cell transplantation for aplastic anemia during January 2014 to October 2016 were analyzed retrospectively. There were 7 males and 8 females, aged from 3 to15 years old, with the median age of 7 years old. Ten cases presented dizziness and headache while other 4 cases presented blurred vision, blind and gaze. Only one case suffered from seizure and loss of consciousness. MRI patterns including distribution, morphology and signal intensity were analyzed after treatment. Follow up MRI were performed after reducing drug dose and symptom remission. The duration of immunosuppressive drugs associated encephalopathy of the 15 cases were 1-14 months, with 6 months in 9 cases. Results Focal lesions were found in 11 cases, in which the deep nuclei were involved in one case and the white matter was involved in 10. Four patients showed both cerebral cortex and white matter lesions, including cerebellum and brainstem invasion in one patient. No corpus callosum lesions were found. Various degree of brain atrophy was found in all patients. Cortical lesions showed swelling and involved subcortical white matter presented as arc shape or strip-like lesions. Patchy patterns were found in deep white matter. Thin layer shaped lesions were found in the periventricular white matter. Small flake-like lesions were found in the brain stem and the cerebellum. The lesions showed hypointensity on T1WI, equal or high signal on T2WI. T2WI FLAIR showed equal or high signal;DWI in the cortex and subcortical white matter lesions showed iso-or high signal, while other lesions were isointense. Eight cases acquired clinical relief in short term without obvious improvement on MRI image. Both clinical symptoms and imaging findings improved in 6 cases. One case showed clinical relief but progression on MRI. Conclusions MRI is an effective way to find immunosuppressive drugs-related encephalopathy in children with aplastic anemia after allogeneic hematopoietic stem cell transplantation. It can help the diagnosis and provide the information for clinical treatment.
6.MRI manifestations of immunosuppressive drugs associated encephalopathy after allogeneic hematopoietic stem cell transplantation in children with aplastic anemia
Yuzhu SHI ; Xinyu YUAN ; Yuchun YAN ; Jie SONG ; Chunli WANG ; Yanmin LUO ; Jiaman WEI
Chinese Journal of Radiology 2017;51(10):782-785
Objective To summarize the MRI manifestations of immunosuppressive drugs associated encephalopathy after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with aplastic anemia. Methods Fifteen patients with immunosuppressive drugs associated encephalopathy after allogeneic hematopoietic stem cell transplantation for aplastic anemia during January 2014 to October 2016 were analyzed retrospectively. There were 7 males and 8 females, aged from 3 to15 years old, with the median age of 7 years old. Ten cases presented dizziness and headache while other 4 cases presented blurred vision, blind and gaze. Only one case suffered from seizure and loss of consciousness. MRI patterns including distribution, morphology and signal intensity were analyzed after treatment. Follow up MRI were performed after reducing drug dose and symptom remission. The duration of immunosuppressive drugs associated encephalopathy of the 15 cases were 1-14 months, with 6 months in 9 cases. Results Focal lesions were found in 11 cases, in which the deep nuclei were involved in one case and the white matter was involved in 10. Four patients showed both cerebral cortex and white matter lesions, including cerebellum and brainstem invasion in one patient. No corpus callosum lesions were found. Various degree of brain atrophy was found in all patients. Cortical lesions showed swelling and involved subcortical white matter presented as arc shape or strip-like lesions. Patchy patterns were found in deep white matter. Thin layer shaped lesions were found in the periventricular white matter. Small flake-like lesions were found in the brain stem and the cerebellum. The lesions showed hypointensity on T1WI, equal or high signal on T2WI. T2WI FLAIR showed equal or high signal;DWI in the cortex and subcortical white matter lesions showed iso-or high signal, while other lesions were isointense. Eight cases acquired clinical relief in short term without obvious improvement on MRI image. Both clinical symptoms and imaging findings improved in 6 cases. One case showed clinical relief but progression on MRI. Conclusions MRI is an effective way to find immunosuppressive drugs-related encephalopathy in children with aplastic anemia after allogeneic hematopoietic stem cell transplantation. It can help the diagnosis and provide the information for clinical treatment.
7.Effect of Chinese medicine on tumor necrosis factor-αin treatment of ul-cerative colitis
Yiyong SONG ; Yi REN ; Yuzhu LIU ; Wenjuan LIU ; Feng LIU
China Modern Doctor 2015;(14):81-83
Objective To assay tumor necrosis factor α(TNF-α) changes during the treatment of ulcerative colitis with the Chinese herbal prescription and study the mechanism of herbal prescription in ulcerative colitis treatment. Meth-ods 60 ulcerative colitis patients were treated with self-made enema treatment, evaluated clinical efficacy of herbal en-ema for ulcerative colitis and detected TNF-α changes in serum. Results After a course of herbal enema treatment, the total effective rate was 95.00% and the level of TNF-α decreased to (20.7±6.6) pg/mL on average. Conclusion Serum tumor necrosis factor level is significantly reduced after the herbal enema treatment. The treatment can enhance immune system function of ulcerative colitis patients and get better clinical efficacy.
8.Regulatory effect of microRNA-126 on macrophage proliferation caused by high glucose stimulation
Yuzhu MU ; Jiayin DENG ; Jiajun LI ; Liting SONG ; Shaoyun JIANG
Chinese Journal of Stomatology 2020;55(12):969-975
Objective:To explore the effects of microRNA-126 (miR-126) on the proliferation of human myeloid leukemia mononuclear cells (THP-1)-derived macrophages in high glucose environment and the regulatory role of miR-126 in periodontitis with diabetes.Methods:THP-1 cells were cultured in vitro and 5 μg/L phorbol-12-myristate-13-acetate was applied to induce THP-1 cells differentiating into macrophages for 48 h in low glucose culture medium (5.5 mmol/L). THP-1-derived macrophages were then cultured with low glucose, medium glucose (15 mmol/L) or high glucose (25 mmol/L) media respectively. The proliferation of THP-1-derived macrophages was detected by cell counting kit-8 (CCK-8) method and the expressions of miR-126 and proliferation-associated factors were detected by quantitative real time PCR (qRT-PCR). The miR-126 mimic or inhibitor was transfected into THP-1-derived macrophages for 72 h. The proliferation of cells was detected by CCK-8 method and the expressions of miR-126 or proliferation-associated factors were detected by qRT-PCR. Results:Increasing glucose concentration decreased the proliferation of THP-1-derived macrophages (day 7, A values in low, medium and high glucose groups were 0.369±0.014, 0.214±0.009 and 0.200±0.010, respectively, P<0.01) as well as the survival rate ( P<0.05), promoted the expression of miR-126, B-cell lymphoma-2 (Bcl-2)-associated X protein (BAX) and caspase-3 ( P<0.05), and suppressed Bcl-2, phosphoinositol-3 kinase regulatory subunit 2 (PIK3R2) expression ( P<0.05). After the miR-126 mimic was transfected in cells in low glucose medium for 72 h, compared with negative control (1.005±0.118), the expression of miR-126 significantly increased (2 980.227±170.431, P<0.05), and the proliferation of THP-1 derived macrophages decreased (negative control: 1.816±0.013, mimic group: 1.310±0.048, P<0.01), the level of BAX and caspase-3 significantly increased ( P<0.01, P<0.05), PIK3R2 and Bcl-2 significantly decreased ( P<0.05, P<0.01). After the miR-126 inhibitor was transfected in cells cultured in high glucose medium for 72 h, compared with negative control (0.723±0.133), the proliferation of inhibitor group increased (0.984±0.049, P<0.05), the level of BAX and caspase-3 significantly decreased ( P<0.01, P<0.05), PIK3R2 and Bcl-2 significantly increased ( P<0.01, P<0.05). Conclusions:High glucose condition can inhibit the proliferation of THP-1-derived macrophages and increase the expression of miR-126. MiR-126 can inhibit the proliferation of THP-1-derived macrophages in high glucose environment through up-regulating the expression of BAX and caspase-3 and down-regulating the expression of PIK3R2 and Bcl-2.
9.Sequence analysis of Pre-S gene in asymptomatic chronic HBV carriers with low-level HBsAg
Tong WANG ; Rui SONG ; Qian LI ; Yuzhu DAI ; Feihu CHE ; Huajun ZHOU ; Changgui SUN ; Jun CHENG
Chinese Journal of Clinical Infectious Diseases 2019;12(2):93-100
Objective To analyze the sequence of Pre-S gene in asymptomatic chronic HBV carriers (ASCs) with low-level HBsAg.Methods The serum samples were collected from 654 ASCs in the First Affiliated Hospital of Zhejiang University School of Medicine , Hangzhou Sixth People’s Hospital and the 903th Hospital of PLA.According to the level of HBsAg , ASCs were divided into low-level HBsAg group (≤10 IU/mL ) and high-level HBsAg group (>10 IU/mL ).The pre-S/S gene amplification and sequencing were performed in 138 ASCs with low-level HBsAg and 100 age-matched ASCs with high-level HBsAg.A phylogenetic tree was constructed to determine the genotype , based on the successful sequencing results of Pre-S gene in the high level HBsAg group , the Pre-S gene reference sequences of the main ASCs genotypes in Eastern China were established.The sequence of Pre-S gene in low-level HBsAg group was analyzed and compared with the reference sequences.SPSS 12.01 statistical software was used to analyze the data.Results Sixty-three cases of Pre-S/S were successfully sequenced in 138 ASCs of low-level HBsAg group, including 52 cases of B genotype and 11 cases of C genotype.Among the 100 cases of high-level HBsAg group, 94 cases of Pre-S/S were successfully sequenced , including 48 cases of B genotype and 46 cases of C genotype.The sequence analysis indicated that in the B genotype , 81 amino acid mutation sites were found in the Pre-S protein of the low-level HBsAg group, including 4 significant mutations: F56I/V, T76A/N/P in the Pre-S1 region, P15L/S/T and Y21T/F/H/N in the Pre-S2 region; while 47 amino acid mutation sites were found in Pre-S protein of high-level HBsAg group, including 3 significant mutations :L34F, V49A and P59S/L in Pre-S1 region.The total number of amino acid mutation sites in the low-level HBsAg group of B genotype was higher than that of the high-level HBsAg group (χ2 =14.008, P<0.05). In the C genotype, 19 amino acid mutation sites were found in the Pre-S protein of the low-level HBsAg group, including 3 significant mutations : W66V/G and A79V in the Pre-S1 region,V32A in the Pre-S2 region; while 39 amino acid mutation sites were found in Pre-S protein of the high-level HBsAg group, including 2 significant mutations: A79V in Pre-S1 region and T49I in Pre-S2 region.The total number of amino acid mutation sites of Pre-S protein in the C genotype was significantly different between the two groups (χ2 =7.571, P<0.05).Conclusion Significant mutations in Pre-S gene may be associated with the persistent expression of low-level HBsAg in ASCs.
10.Function of the IL28B gene in Hepatitis B virus infection.
Chenglin ZHANG ; Yuzhu SONG ; Yumei MA ; A'mei ZHANG
Chinese Journal of Biotechnology 2017;33(2):187-195
Hepatitis B virus (HBV) that causes Hepatitis B with a high chronic rate, could lead to hepatic cirrhosis and hepatocellular carcinoma. The IL28B gene belongs to a new interferon family λ and its genetic polymorphisms are identified to be associated with treatment effect and viral clearance. The purpose of this review is to discuss the role of the IL28B gene in treatment response and viral clearance of HBV-infected individuals, and further to reveal the mechanisms. This review could provide a theoretical basis for personalized medicines of HBV patients.