1.Effect of miR-29b transfected by anti-TGF-βⅡR ScFv/Ck/tP fusion protein on liver fibrosis
Fujun YU ; Jianjian ZHENG ; Shenmeng GAO ; Peihong DONG
Journal of Chinese Physician 2012;(10):1331-1334
Objective To observe the transfection efficiency and anti-fibrotic effect of miR-29b transfected by anti-TGF-β Ⅱ R ScFv/Ck/tP fusion protein (new vector) in hepatic stellate cell (HSC),and to provide a new vector in gene therapy for liver fibrosis.Methods The liposome vector,new vector,and lentiviral vector were used as transfection reagents to transfect miR-29b into HSC.Transfection efficiency was observed under fluorescence microscope and flow cytometry.Collagen α1 (Ⅰ) mRNA and protein expression in different groups were analyzed by real-time RT-PCR and Western Blot,respectively.Results Compared to the control,transfection efficiencies in lentiviral vector,new vector,and liposome vector groups were about 70%,58%,and 29%,respectively.Collagen α1 (Ⅰ) mRNA expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 70%,50%,and 38%,respectively ((t =6.316,P <0.01 ; t =4.082,P <0.01 ; t =3.014,P <0.05).Collagen α1(Ⅰ) protein expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 59%,41%,and 27%,respectively (t =4.209,P <0.01; t =4.033,P <0.01; t =2.842,P <0.05).Conclusions The new vector constructed by us has a high transfection efficiency.MiR-29b transfected by the new vector has a good anti-liver fibrosis effect.
2.Effects of ethyl pyruvate pretreatment on liver injury in rats undergoing orthotopic liver transplantation
Jianjian YU ; Hongyin DU ; Wenli YU ; Yiqi WENG ; Yuliang WANG ; Weihua LIU
Chinese Journal of Anesthesiology 2012;32(3):304-306
Objective To investigate the effects of ethyl pyruvate (EP) pretreatment on the liver injury in rats undergoing orthotopic liver transplantation (OLT).Methods Forty male adult SD rats,weighing 220-250 g,served as liver transplant donors and recipients.The recipient rats were randomly divided into 3 groups ( n =8 each):sham operation group (group S),OLT group and EP group.Group S only underwent simple laparotomy.The model of OLT was established according to the modified Kamada's two-cuff technique in groups OLT and EP.EP 40 mg/kg was injected via the caudal vein at 1 h before skin incision in group EP.Venous blood samples were taken at 2 h of neohepatic phase to determine the serum activities of alanine amino-transferase (ALT) and aspartate amino-transferase (AST).Hepatic specimens were obtained to detect the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD).Results Compared with group S,the levels of serum ALT and AST and MDA were significantly increased and the activity of SOD was significantly decreased in groups OLT and EP ( P <0.05 or 0.01).The levels of serum ALT and AST and MDA were significantly lower and the activity of SOD was significantly higher in group EP than in group OLT ( P < 0.05 or 0.01 ).Conclusion Ethyl pyruvate pretreatment can attenuate the liver injury in rats undergoing OLT.
3.Diagnostic and therapeutic status of the elderly outpatients with benign prostatic hyperplasia in Guangzhou
Wenke HAO ; Yunjuan QIAN ; Xilin ZHANG ; Dongmei CAI ; Jianjian YAO ; Feng YU ; Jing CHEN
Chinese Journal of Geriatrics 2010;29(11):949-951
Objective To evaluate the diagnostic and therapeutic status of lower urinary tract symptoms (LUTS) in the elderly outpatients with benign prostatic hyperplasia (BPH) in Guangzhou.Methods The investigation was conducted in outpatient departments for elderly patients in 3 hospitals of Guangzhou. The eligible male patients with LUTS/BPH were chosen from patients who came to the clinic in the process of investigation. Results The 29.7% (1824/6140) of them had history of LUTS/BPH previously or presently. In 134 patients received medical questionnaire, the proportion of LUTS/BPH patients with mild, middle and severe degree based on international prostate symptom score (IPSS) was 24.5%, 72.5% and 3.0% respectively. The abnormal percentage of PSA was 37. 3 %. The most common examinations that patients received were directeral rectun examination (96.8%), PSA (88. 7%) and transabdominal ultrasonography (84.8%). The 44.7% of cases were given the treatment of 5α-reductase inhibitor, and 24.7% of cases received a combination therapy of 5α-reductase inhibitor plus α receptor blocker. The 16.7% of cases were given a combination treatment of 5α-reductase inhibitor plus herbs. The proportion of therapy with α receptor blocker and herbs was similar (6.8% and 7.1% respectively). Conclusions LUTS/BPH is one of the most common diseases in elderly male patients. The examinations conducted by doctors are unreasonable.The doctors are easy to ignore inquirement of patient's history and IPSS. The drugs conducted by doctors are reasonable basically.
4.Effects of hydrogen-rich kidney preservation solution on renal cold ischemia/reperfusion injury in rats
Hengchang REN ; Hongyin DU ; Wenli YU ; Yiqi WENG ; Yongwang WANG ; Gang WANG ; Rubin XU ; Jianjian YU ; Yuliang WANG
Chinese Journal of Anesthesiology 2012;(12):1491-1494
Objective To investigate the effects of hydrogen-rich HC-A solution,the self-made kidney preservation solution,on renal cold ischemia/reperfusion (I/R) injury in rats.Methods Twenty-four healthy male Wistar rats,aged 8-10 weeks,weighing 200-250 g,were randomly divided into 3 groups (n =8 each):control group (H1 group),common kidney preservation solution group (H2 group) and hydrogen-rich HC-A kidney preservation solution group (H3 group).In H1 group,only the right kidney was removed.In H2 group,the left kidney was perfused with and cold stored in 4 ℃ common HC-A kidney preservation solution.In H3 group,the left kidney was perfused with and cold stored in a container filled with 4 ℃ common HC-A kidney preservation solution.Blood samples were obtained from the inferior vena cava at 24 h of reperfusion to detect the levels of serum blood urea nitrogen (BUN),creatinine (Cr),TNF-α and IL-6.Left kidneys were removed for determination of malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) contents and for examination of the pathological changes in renal tissues (by light microscopy).Results The levels of serum BUN,Cr,TNF-α and IL-6 and contents of MDA and 8-OHdG were significantly higher in H2 and H3 groups than in H1 group,and lower in H3 group than in H2 group (P < 0.05).There were no significant pathological changes in renal tissues in H1 group,the damage to renal tubules was obvious in H2 group and the damage to renal tubules was significantly ameliorated in H3 group as compared with H2 group.Conclusion Hydrogen-rich kidney preservation solution can attenuate renal cold I/R injury in rats.
5.The molecular regulatory mechanism of co-treatment with LA and PCA on P38 MAPK signaling pathway in the neurons of TX suckling mice
Chenchen XU ; Jianjian DONG ; Nan CHENG ; Xun WANG ; Xuen YU
Chinese Journal of Nervous and Mental Diseases 2017;43(8):490-495
Objective To detect the molecular regulatory mechanism of co-treatment with LA and PCA on P38 MAPK signaling Pathway in the Neurons of Wilson's Disease Model-TX mice.Methods The neurons of TX suckling mice were isolated and cultured by primary method,and were divided into control group,model group,ALA group,PCA group and combined group.Flow cytometry was used to analyze the expression of ROS and JC-1.Western blot was used to detect the expression of P38 MAPK,Cyt C,Caspase 9 and Caspase 3.Results Flow cytometry results showed that MFI of ROS was 59.29±1.22,53.19±1.34 and 52.46±1.23 in ALA,PCA and co-treatment.ALA,PCA and co-treatment could significantly reduce the release of ROS and enhance the fluorescence intensity of JC-1 (P<0.01).Compared with ALA group and PCA group,combined group could reduce the release of ROS and significantly enhance the fluorescence intensity of JC-1.Western blot indicated that the expression levels of P38 MAPK,Cyt C,Caspase 9,Caspase 3 in the neurons of model group had a remarkable increase compared with control group.Compared with the model group,the three treatment groups could decrease the expression levels of P38 MAPK,Cyt C,Caspase 9 and Caspase 3 in the neurons of TX suckling mice (P<0.01).Meanwhile,the protein levels of P38 MAPK,Cyt C,Caspase 9 and Caspase 3 had a significant decrease compared with ALA group and PCA group.Conclusion he present findings suggest that co-treatment with LA and PCA can increase the copper excretion,reduce copper-induced mitochondria damage and attenuate the neurotoxicity,which in turn decrease neuronal apoptosis and improve neurological impairment of WD.
6.Mechanism of Traditional Chinese Medicine in Prevention and Treatment of Parkinson's Disease:A Review
Xiaolong SHANG ; Yu WANG ; Chenchen XU ; Jianjian DONG ; Wenlong AI ; Liangjie ZHANG ; Chunling CI ; Xun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(17):248-259
Parkinson's disease (PD) is a progressive chronic neurodegenerative disorder with a complex pathogenesis involving oxidative stress, neuroinflammation, mitochondrial dysfunction, and other factors. Currently, the clinical treatment of PD mainly includes levodopa, dopamine receptor agonists, monoamine oxidase B inhibitors, catechol-O-methyltransferase inhibitors, and anticholinergic drugs, but there is a lack of disease-modif g therapies that can definitively improve disease progression. According to the understanding of traditional Chinese medicine (TCM), PD is characterized by asthenia in origin and sthenia in superficiality. It is primarily caused by liver-kidney Yin deficiency, Qi-blood insufficiency, and closely related to wind, fire, phlegm, and blood stasis. Numerous clinical practices have shown that TCM has significant clinical value in the prevention and treatment of PD, the management of motor and non-motor symptoms, and the neuroprotection of dopaminergic neurons. The underlying mechanisms of TCM include antioxidative stress, anti-neuroinflammation, and regulation of mitochondrial dysfunction. This article categorized and summarized the pathogenesis of PD, systematically elucidated the pharmacological actions and molecular mechanisms of TCM monomer extracts and compounds in the prevention and treatment of PD, and provided the latest clinical research progress, aiming to provide references for the development and clinical use of TCM for PD.
7.False-positive results of rifampicin resistance in Xpert MTB/RIF testing of samples with extremely low bacterial loads
Youyi RAO ; Chang LIU ; Qiudan XIN ; Jianjian GUO ; Jian YU ; Jun CHEN
Journal of Public Health and Preventive Medicine 2024;35(5):24-27
Objective To investigate the causes of false-positive rifampicin resistant results in Xpert MTB/RIF (Xpert) test for samples with extremely low bacterial loads. Methods A total of 346 samples with extremely low bacterial loads and rifampicin-resistance results from Wuhan Pulmonary Hospital between June 2017 and March 2021 were collected. The samples were divided into probe-delayed and probe dropout groups based on amplification results. Mycobacterial culture and proportion method drug susceptibility testing were performed, and Xpert retesting was conducted for strains with discordant drug susceptibility results. Results Out of the 346 samples, 195 samples (56.36%) were positive in culture. Upon Xpert retesting, among the 64 Xpert-resistant but proportion method-sensitive strains, the proportions of samples in the delayed probe group with mutations in the probe D and probe E were 4.55%(1/22) and 13.33%(2/15), respectively. In the probe dropout group, the proportions of samples with mutations in the probe A and probe E were 75.00% (9/12) and 80.00% (8/10), respectively. The false-positive rifampicin resistance rates in the delayed probe and probe dropout groups were 78.26% (36/46) and 3.36% (5/149), respectively. Conclusion The main reasons for false-positive rifampicin resistance results in the Xpert test for samples with extremely low bacterial loads were probe delay in the D and E probes, followed by low-level drug-resistant mutations in the A and E probes.