1.Expression of acid sensing ion channel 3 in the lung tissue of rats with, acute lung injury
Lan SHAO ; Yansheng CHEN ; Shaoqun XU ; Haibo QIU
Chinese Journal of Emergency Medicine 2009;18(5):466-470
Objective To explore the expression of acid seining ion channels-3 (ASIC3) in lung tissue in rats with acute lung injury (ALI). Method Twenty four male Sprague-Dawley (SD) rats were randomly divided into four groups: LPS groups (LPS 2 h, LPS 4 h, LPS 6 h group, n=6), stimulated by LPS for 2, 4, 6 hours, respectively; normal control group, injected with saline (NS group, n=6). The ALI models were produced through venous injection of LPS, and the criteria was the characteristic pathological changes in the lung tissue. Ar-terial blood gas analysis was observed, lung wet and dry weight ratio (W/D), lung histopathology and ASIC3 ex-pression were detected. Data were expressed as mean±standard deviation. Independent Sample T test and One-way ANOVA and Kendall's tau_b were used for comparison in SPSS 13.0, and changes were considered as statistieal-ly significant if P value was less than 0.05. Results The partial pressure of oxygen (PaO2) in LPS 2 h, LPS 4 h, LPS 6 h group was (67.47±6.01), (59.17±7.18), (52.54±7.62) , respecively, and was significantly lower than that in eontrol group (98.15±1.06) (P<0.01). Compare with control group, pH was significantly lower in LPS4 h group (7.28±0.04), LPS6 h group (7.24±0.03) (P<0.01). Inflammation cells gradually increased, alveolar septum was widened, edema existed in interstitial spaces, and pulmonary structures gradually destroyed in LPS groups.The expression of ASIC3 in LPS4 h, LPS 6 h group was (205.91±10.12), (196.51± 18.60), respectively, and was significantly lower thanthat in control group (220.23±10.11) (P<0.05). The W/D in LPS 6 h group was (5.18±0.21), and was significantly higher than that in the control group (4.45± 0.18) (P<0.05). Conclusions ASIC3 is expressed in alveolar epithelial cells and bronchial epithehal cells in LPS-induced ALI rats.
2.Effects of 137 Cs γ-rays on proliferation, differentiation and mineralization of osteoblastic cells in vitro
Jing QIU ; Guoying ZHU ; Shuzhu GU ; Xiao CHEN ; Chunlin SHAO
Chinese Journal of Radiological Medicine and Protection 2012;32(2):191-195,203
Objective To evaluate the effect of gamma irradiation on the proliferation,differentiation,and mineralization of murine osteoblastic cells,and to investigate the related molecular mechanism.Methods Osteoblastic cells were irradiated by different doses (0,0.5,1.0,2.0,5.0 Gy)of 137Cs γ-rays.Cell morphology was observed with a microscopy,cell viability was analyzed by MTT assay,and ALP activity was analyzed by the methods of enzyme histochemistry and PNPP.Meanwhile,gene expressions of ALP,osteocalcin (OC),collagen Ⅰ,osteoprotegerin (OPG) and receptor activator of nuclear factor-kB ligand (RANKL) were measured by semi-quantified RT-PCR.Results Cell viability decreased with the radiation doses over 1.0 Gy ( t =6.197 - 18.677,P < 0.05 ).After radiation with a dose over 2.0 Gy,the cell number and the junctions of cell protrusions decreased,the cells had low refractivity and the activity and mineralization ability of ALP were also inhibited ( t =2.790 -2l.374,P <0.05).In addition,the expressions of ALP and OC mRNA were down-regulated significantly (t =3.563 -16.508,P < 0.05) when the radiation dose was higher than 0.5 Gy,and the expressions of OPG,OPG/RANKL mRNA were down-regulated ( t =12.942,4.954,P < 0.05 ) at 5 Gy.But the expressions of collagen Ⅰ and RANKL mRNA were not affected by irradiation.Conclusions The osteoblastic cells were significantly influenced by γ-irradiation,including morphological changes,inhibition of cell proliferation,differentiation and mineralization ability. Meanwhile,mRNA expressions of ALP and OC were downregulated.OPG/RANKL may be a main pathway of osteoblastic cell damage under high dose radiation.
3.Effect of Qiling Decoction combined HAART on expression levels of Treg cells and Th17 in HIV/AIDS patients.
Wen-Fang XU ; Yong WU ; Guo-Shao PAN ; Jian-Ping ZHONG ; Shao-Bo LAN ; Xue-Fang CHEN ; Qiu-Qiong LU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(2):157-161
OBJECTIVETo explore the effect of Qiling Decoction (QD) combined highly active antiretroviral treatment (HAART) on expression levels of peripheral blood Th17 and Treg cells in HIV/AIDS patients.
METHODSTotally 55 HIV/AIDS patients were randomly assigned to the treatment group (28 cases) and the combination group (27 cases). Besides, 21 HIV negative patients were recruited as the healthy control group. Those in the treatment group received HARRT alone, while those in the combination group received HAART combined QD. The observation lasted for 24 weeks. Meanwhile, according to peripheral blood CD4+ T cell counts before treatment, HIV/AIDS patients were assigned to three subgroups. For patients in subgroup 1, 1 cells/microL < CD4+ T cell counts < or = 100 cells/microL; For patients in subgroup 2, 101 cells/microL < CD4+ T cell counts < or = 200 cells/lL; For patients in subgroup 3, 201 cells/microL < CD4+ T cell counts < or = 350 cells/microL. Expression of peripheral blood Th17 and Treg cells, and number of CD4+ T cell counts were detected using flow cytometry (FCM)in HIV/AIDS patients at the pre-treatment baseline, week 4, 12, and 24, as well as those in the healthy control group.
RESULTSCompared with the healthy control group, CD4+ T cell counts and the baseline expression level of Th17 cells in the peripheral blood of HIV/AIDS patients significantly decreased, the expression level of Treg cells significantly increased P < 0.01). Compared with before treatment in the same group, CD4+ T cell counts all increased at week 4, 12, and 24 in the two treatment groups, showing statistical difference (P < 0.05, P < 0.01). There was no statistical difference in the effective rate at various CD4+ T cell levels between the two groups (P > 0.05). There was no statistical difference in expression levels of Th17 and Treg cells between the combination group and the treatment group at any time point (all P >0.05). The Th17/Treg ration significantly increased in the combination group after 24 weeks of treatment, showing statistical difference when compared with the treatment group (U = 2.135, P = 0.038).
CONCLUSIONQD could improve the immune balance of Th17/Treg cells, which might be one of its mechanisms for improving HIV/AIDS patients' immunity.
Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology
5.Hemodialysis membrane:its biocompatibility and application in severe pancreatitis
Ou CHEN ; Xiong QIU ; Zeyong SHAO ; Wenjie LUO ; Changping LI ; Muhan LV
Chinese Journal of Tissue Engineering Research 2016;20(8):1177-1182
BACKGROUND: Continuous blood purification can remove cytokines and inflammatory mediators, maintain homeostasis and prevent the occurrence of multiple organ dysfunction syndrome in patients with severe pancreatitis, which has become the main therapy for severe pancreatitis. Since the hemodialysis technology began to be applied clinical y, the biological and physicochemical properties of hemodialysis membrane materials have been studied. A variety of hemodialysis membranes have been developed in order to improve the biocompatibility and anticoagulant effect in vitro. OBJECTIVE: To investigate the application effect of hemodialysis membranes on severe pancreatitis. METHODS: Ten Wistar rats were selected to make rat models of severe pancreatitis and then were randomized into two groups (n=5 per group): homophone membrane group and polysulfone membrane group. Hemodialysis- related biochemical parameters were detected in the two groups. RESULTS AND CONCLUSION: Compared with the homophone membrane, ultrafiltration coefficient, creatinine clearance, blood urea nitrogen clearance, phosphorus clearance, number of circulating endothelial cel s, and levels of plasma nitric oxide and asymmetric dimethylarginine were significantly lower in the polysulfone membrane group (P < 0.05). Vitamin B12 clearance and amount of pre-congestion increased significantly in the polysulfone membrane group as compared with the homophone membrane (P < 0.05). These findings indicate that the polysulfone membrane for hemodialysis has good biocompatibility, and keeps a stable environment in vivo for severe pancreatitis patients.
6.Preliminary efficacy of bevacizumab for cerebral radiation necrosis
Mianshun PAN ; Yong LI ; Shujun QIU ; Lei CHEN ; Xianjun SHAO ; Li ZHANG ; Guoyu ZHANG ; Fenghua ZHUGE
Chinese Journal of Radiation Oncology 2015;(4):434-437
Objective To evaluate the preliminary clinical efficacy of bevacizumab for cerebral radiation necrosis (CRN). Methods Nineteen patients with CRN for whom the treatment with steroids and mannitol failed were retrospectively analyzed with a total of 22 lesions. Except for 5 lesions confirmed by pathological evidence, all lesions were confirmed by the following imaging evidence:1. computed tomography (CT)?or magnetic resonance imaging (MRI)?enhanced lesions showed loss of tension and were accompanied by substantial edema;2. CT?or MRI?enhanced lesions had a low perfusion pressure;3. magnetic resonance spectroscopy indicated that the enhanced areas had a decreased choline peak; 4. positron emission tomography showed that the fluorodeoxyglucose uptake was substantially reduced in the enhanced areas. All patients were given 5 mg/ kg bevacizumab at an interval of 14 days for 2?6 cycles. MRI examination was performed in each cycle before treatment, and the enhanced lesions on T1?weighted images ( T1 WI) and edema on T2?weighted images (T2 WI) were compared before and after treatment. The clinical symptoms, Karnofsky Performance Status ( KPS), and adverse reactions in all patients were evaluated. Comparison before and after treatment was performed by paired t test. Results All 19 patients completed the treatment successfully and there were no severe adverse reactions. The clinical symptoms of patients were substantially improved after the second cycle of treatment, and the KPS score increased by 26?? 8 on average. The visible volume of enhanced lesions on MRI T1 WI was significantly reduced by 54?? 8% after treatment (P= 0?? 000), while the visible volume of edema on MRI T2 WI was reduced by 80?? 7% after treatment (P= 0?? 000). The follow?up time ranged from 3 to 12 months with a mean value of 5?? 6 months. Eleven patients kept clinical improvement in CRN, four patients had recurrence, and four patients died from tumor progression. Conclusions Bevacizumab is preliminarily confirmed to substantially improve the clinical symptoms and quality of life in patients with CRN.
8.Urine metabonomic study on long-term use of total ginsenosides in rats.
Xie XIE ; Shao-Qiu CHEN ; Ying-Fang LV ; Xiao-Yan WANG ; Wei JIA
China Journal of Chinese Materia Medica 2014;39(23):4675-4679
Due to its effect of systems regulation and promotion on body, Ginseng is always referred to be long-term used as a dietary supplement. But it was still unclear about its target of the tonic effects and also the side-effects long-term use may bring. Urine metabolomic method is suitable for long-term studies of pharmaco-dynamics, pharmacology and toxicology of traditional Chinese medicine because of its characteristics of non-invasive and monitoring the whole-body metabolism. This study was designed to detect the dynamic variation of rat urine metabolome along with a long-term administration of total ginsenosides using GC-TOF based metabolomic technology. Our result showed that either short-term or chronic administration of ginsenosides did not impact the rat urine metabolome significantly (as the PCA subgroup was not successful). By comparison, the short-term (1-3 w) dose of ginsenosides had the biggest metabolic influence including TCA cycle, catecholamines and neurotransmitter amino acids. Medium-term (6-10 w) dose had a gradually lower effect and long-term (27 w) dose almost had no effect. Our study indicates that both short and long-term administration of ginsenosides showed almost no obvious side-effect on the experimental animals.
Animals
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Drugs, Chinese Herbal
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metabolism
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Ginsenosides
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metabolism
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urine
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Male
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Metabolomics
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Panax
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metabolism
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Rats
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Rats, Wistar
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Time Factors
9.Two-Step MS-PCR Combined With ELISA Method for the Detection of Drug Resistance Mutations in HIV-1 RT Gene
Hong-Qiu HE ; Shao-Hui CHENG ; Bin LIU ; Wei-Zu CHEN ; Cun-Xin WANG ;
China Biotechnology 2006;0(08):-
Highly Active Anti-Retroviral Therapy (HAART) has effectively inhibited the prevalence of HIV-1 and reduced the death rate caused by AIDS. In recent years,the emergence of resistance-conferring RT gene mutations in HIV-1 strains has become the major reason for HAART failure. The detection of drug resistance is important for the HAART regimen choice and novel drug development. A novel assay for the detection of HIV-1 RT drug resistance mutations was developed. HIV-1 drug resistance and wild strains in B subtypes were investigated using Two-Step Mutagenically-Separated PCR (MS-PCR),and point mutations including M41L,K70R,K103N,Y181C,T215F were detected. A longer mutant type primer was designed,using microplates hybridization and ELISA technique to detect several point mutations within a mixed mutant-wild type population. The results indicate that the Two-Step MS-PCR is as sensitive and specific as that in the traditional MS-PCR and MS-PCR combined with ELISA can give a good P/N quotient with better sensitivity,low cost,relatively less time consumption and high-throughput screening. It will be used in clinic usage for the detection of HIV-1 drug resistance mutations as well as other point mutations.
10.Prokaryotic Expression and Functional Study of HIV-1 Envelope Glycoprotein gp41 Helical Bundle
Bin LIU ; Hong-Qiu HE ; Shao-Hui CHENG ; Wei-Zu CHEN ; Cun-Xin WANG ;
China Biotechnology 2006;0(07):-
HIV-1 envelope glycoprotein gp41,which is a hopeful target for HIV-1 fusion inhibitors,plays a critical role in the fusion of viral and cellular membranes.In order to build up the screening assay of HIV-1 fusion inhibitors targeting gp41,HIV-1 gp41 5-helix and 6-helix were expressed in prokaryotic cells.Gp41 5-helix and 6-helix recombined plasmids were constructed by using PCR,enzyme digestion and ligation taking the clade B HIV-1 genome as a template.The plasmid was transferred into E.coli BL21(DE3)and then induced by IPTG.The expressed protein was purified by affinity chromatography after denaturation and renaturation.The SDS-PAGE analysis was used during expression and purification.Native-PAGE was used to identify the interaction between gp41 5-helix and T-20.The result will be helpful to build up the screening assay of HIV-1 fusion inhibitors targeting gp41.