1.Progress in microbiome and its application to pharmacological and toxicological research of traditional Chinese materia medica
Hong DENG ; Chunqi WU ; Tao JIANG ; Quanjun WANG
Chinese Journal of Pharmacology and Toxicology 2016;30(9):975-982
Microbiome is a novel research field related to human health,agriculture,bio-energy and the environment. Gut microbiome has received much attention from researchers recently. Studies have shown that gut flora is related to some diseases,such as digestive disease(inflammatory bowel disease),metabolic disease(type 2 diabetes), cardie-cerebral vascular disease(Parkinson disease). Traditional Chinese materia medica(TCMM) has long been used as a tonic and taken in a large amount. Gut flora has an effect on pharmacology and toxicology of TCMM after entering the gastroin?testinal tract. This article is intended to review recent researches on microbiome,common detection techniques and the relationship with hepatotoxicity induced by Polygonum multiflorum Thunb.,scutel?laria baicalensis that directly affects the intestinal tract,nephrotoxicity induced by Rhizoma Alismatis and pneumonia induced by Xiao-Chaihu-Tang.
2.Screening, expression and characterization of single chain Fv (scFv) against basic fibroblast growth factor
Hong WANG ; Jun TAO ; Ning DENG ; Lijun ZHOU ; Junjian XIANG
Chinese Journal of Microbiology and Immunology 2010;30(7):674-677
Objective To obtain the specific human scFv basic fibroblast growth factor(bFGF)using phage antibody library technology. Methods The library was panned with human recombinant bFGF for 4 rounds. The antigen binding activities of random clones were tested by ELISA in order to select specific antibodies, which were then examined by DNA sequence analysis. Results The positive clone selected from the 104 random clones was able to bind bFGF specifically, while not able to bind other growth factors,such as aFGF, VEGF(vascular endothelial growth factor). By competition ELISA assay we found one clone 44 could inhibit bFGF binding to FGFR1. Conclusion Seven specific human phage antibody against bFGF was obtained by phage display technique, one clone could inhibit bFGF binding to its high affinity receptor FGFR1.
3.Production of a dual model probe for photoacoustic imaging and fluorescence imaging targeting integrinαvβ6
Chao ZHANG ; Kai HONG ; Yang YU ; Anyu TAO ; Youbin DENG ; Jie WAN
Chinese Journal of Ultrasonography 2016;(1):81-85
Objective To develop a probe for photoacoustic imaging and fluorescence imaging targeting integrin αvβ6 . Methods The probe was separated by RP‐HPLC .Molecular weight and the maximum absorption wavelength of the probe were detected by mass spectrum instrument and optical spectrum instrument . Various concentrations of the probe were detected by photoacoustic imaging and fluorescence imaging . The stability of the probe was evaluated when exposed under laser . Targeting of the probe on integrinαvβ6 was evaluated in cell uptake assay with integrinαvβ6 positive and negative cells . The minimum number of cells that could be detected by photoacoustic imaging and fluorescence imaging was also evaluated . Results The probe ICG‐peptide was separated from reaction mixture by RP‐HPLC .The probe had a retention time of 21 .4 minutes and m/z of 4 727 . The labeling ratio of the probe was 1∶1 . The maximum absorption wavelength of the probe was 790 nm . The photoacoustic signal was linearly dependent on the concentration of the probe . The fluorescence signal was linearly dependent on the concentration of the probe when the concentration was smaller than 1 .5 × 10 -5 mol/L . The lowest concentration of the probe that could be detected above the background by photoacoustic imaging and fluorescence imaging was 0 .09 × 10-5 mol/L and 0 .05 × 10-5 mol/L ,respectively . No obvious decrease of the photoacoustic signal was observed after the probe was scanned 20 times ( each time lasted for 1 min) by laser . There existed differences ( P <0 .001) in cell uptake of the probe with various concentrations and reaction time between A431 cells (αvβ6 positive) and 293T cells (αvβ6 negative) . Cell uptake was inhibited by the addition of 5μmol/L unlabeled peptide in A431 cells ( P = 0 .001 ) . The lowest number of the labeled A431 cells detected by photoacoustic imaging and fluorescence imaging was 0 .4 × 106 and 0 .05 × 106 ,respectively . Conclusions The dual functional photoacoustic and fluorescence probe targeting integrin αvβ6 was successfully developed . The targeting and sensitivity of the probe makes it potentially useful in early detection of αvβ6 positive tumors .
4.Peripheral blood T lymphocytes cell level of different grades of cerebral gliomas patients before and after operation and its clinical significance
Chunhua XU ; Yue LIU ; Limin XIAO ; Shengze DENG ; Changgui GUO ; Erming ZENG ; Tao HONG
Chongqing Medicine 2016;(2):180-182
Objective To explore the peripheral blood T lymphocytes cell level in different grades of cerebral gliomas pa-tients at the time of before and after operation and its clinical significance .Methods A total of 80 cases of brain tumor patients from February 2010 to February 2012 in this hospital were chosed as study objects ,included 57 cases of glioma ,23 cases of other brain tumors ;57 cases of glioma were divided into 19 cases of low grade group and 38 cases of high grade group accorded to WHO grading standards ,and 50 cases of healthy people in same period were selected as control group .Venous blood in three groups were extracted at 1st day and 1st week after operation ,to detected the level of T lymphocyte subsets in peripheral blood and analyze the relationship between the different grade gliomas and prognosis .Results The peripheral blood CD3 + ,CD4 + ,CD4 + /CD8 + in control group were significantly higher than that of other brain tumor group and the glioma group ,CD8 + was significantly lower than that of the two groups(P< 0 .05) .Compared with other tumor group ,peripheral blood CD3 + ,CD4 + ,CD4 + /CD8 + in glioma group de-creased significantly ,CD8 + increased significantly(P< 0 .05) ;before and after the operation ,the CD3 + and CD4 + /CD8 + levels were significantly higher in low grade group than in the high grade group ,CD8 + was significantly lower in low grade group than in the high grade group(P< 0 .05) ,at the time of 1st week after operation ,CD3 + and CD4 + /CD8 + increased ,CD8 + decreased significant-ly of two groups when intra-group comparison (P < 0 .05) .The median survival time were 31 months in low grade group and 13 months in high grade group ,and the singnificance was found in two groups(P< 0 .05) .The median survival time were 34 months in CD4 + /CD8 + > 1 group and 17 months in CD4 + /CD8 + < 1 group ,the singnificance was found in two groups(P< 0 .05) .Conclusion The immune function of patients with brain glioma is inhibited ,the higher the malignancy of the tumor ,the more obvious of im-mune inhibition ;T lymphocyte subsets level could be used as evaluating index of malignant degree and prognosis of brain glioma .
5.Development of totally enclosed slide culture medium and its application in filamentous fungi culture
ke Dong CHEN ; xing Cun DENG ; tao Hong XU
Chinese Journal of Clinical Laboratory Science 2017;35(10):739-743
Objective To develop a safe,practical,disposable slide culture medium for fungi culture.Methods Based on the principle of traditional slide culture medium with small steel ring,a plate of polyvinyl chloride (PVC) with high transparency was chosen as the bottom material of the new medium which held the size of the conventional slide (25.2 mm × 75.9 mm × 1.05 mm) with frosted design at both ends.The diameter of inoculation hole was 2.4 mm on the rounded culture dish which was designed with diameter of 19.4 mm and side height of 3.75 mm.The users could inject potato dextrose agar (PDA) or other medium into the culture dish as needed and then sealed it with a plug.The upper cover was prepared with toughened glass.The improved disposable slide culture medium should be kept in humidifying box with sponge strips in the water tanks of both sides and sealed with cap.The growth status,microscopic morphology and staining result of filamentous fungi in the totally enclosed slide culture medium were observed and the preservative status of the prepared medium was also monitored simultaneously.Results The effects of the improved slide culture medium were satisfactory in clinical practical application.The growing status of fungi could be observed visually and the pigment was clear.The original growth form of fungi could be monitored under microscope and dye material could be perfused directly to stain with good results.The appearance and the volume of packaged slide medium were unchanged after preservation at 4 ℃ for 3 months.Conclusion An improved slide culture medium was successfully developed,which should be easy to operate,high visible,satisfactory for sealing effects and reliable for culture performance with high biological safety.The growth status of fungi could be observed under microscope at any time,and the medium could also be monitored under oil immersion lens directly and stained with cotton blue.The improved medium could be used in morphological examination for fungi in different levels of medical laboratories since its favorable results in clinical application.
6.Therapeutic effect of collagen from Cyanea nozakii on adjuvant arthritis in rats
Wen-Tao ZHANG ; Lu-Hong TANG ; Wei CHEN ; Dong-Qun CHEN ; Chao DENG ;
Chinese Journal of Marine Drugs 1994;0(04):-
Objective To investigate the suppressive effects of collagen from Cyanea nozakii on adjuvant induced arthritis inrat.Methods Rats with adjuvant arthritis received different do- ses of Cyanea nozakii collagen by intragastric administration for two weeks.Incidence and severity of arthritis were assessed by calculation of mean arthritis index,the concentrations of NO,MDA and activity of SOD in the serum were examined.Results Cyanea nozakii colla- gen at different doses could ameliorate the adjuvant induced arthritis,suppress the concen- trations of NO,MDA and increase the activity of SOD in the serum.Conclusion Cyanea noza- kii collagen has therapeutic efficacy in treatment of adjuvant arthritis rats,and the mecha- nism of Cyanea nozakii collagen is probably related to the antioxidation.
7.Autologous oxygen-delivering biomimetic nanoscaffold composited with chondrocytes reconstructs the temporomandibular joint
Hong WANG ; Tianan LIAO ; Tao WANG ; Liangbin FU ; Guangwei HU ; Wei DENG
Chinese Journal of Tissue Engineering Research 2016;20(30):4469-4475
BACKGROUND:The autologous oxygen-delivering biomimetic nanoscaffold is composited with chondrocytes in simulated microgravity to construct the active tissue-engineered bone, which not only has excel ent osteogenic potential characteristics, but also has autologous releasing oxygen, and additional y can effectively prevent early transplant failure caused by sufficient revascularization and hypoxia. OBJECTIVE:To investigate the effects of autologous oxygen-delivering biomimetic nanoscaffold composited with chondrocytes on repairing different types of temporomandibular joint condylar fractures. METHODS:Total y 30 Sprague-Dawley rats were randomly divided into control group, experimental groups 1 and 2 (n=10 per group). The autologous oxygen-delivering nano-hydroxyapatite/chitosan scaffold composited with chondrocytes were transplanted into rats with temporomandibular joint condylar longitudinal fracture in the experimental group 1 and those with temporomandibular joint condylar transverse fracture in the experimental group 2, respectively. At 1, 3 and 6 weeks after transplantation, the number of proliferating cel nuclear antigen-positive cel s was detected by immunofluorescence assay;the chondrocyte apoptosis was detected using TUNEL method, and expressions of col agen type II, Sox9 and vascular endothelial growth factor were observed by RT-PCR technology. RESULTS AND CONCLUSION:The number of proliferating cel nuclear antigen-positive cel s in the experimental groups 1 and 2 were significantly higher than that in the control group at different time points after transplantation (P<0.05). At 3 and 6 weeks after transplantation, the number of apoptotic chondrocytes in the experimental groups 1 and 2 was significantly lower than that in the control group (P<0.05);significantly higher and highest expressions of col agen type II, Sox9 and vascular endothelial growth factor were found in the experimental groups 2 and 1 compared with the control group, respectively (P<0.05). Additional y, compared with the experimental group 2, the number of proliferating cel nuclear antigen-positive cel s was significantly lower in the experimental group 1 at 3 and 6 weeks after transplantation (P<0.05);the number of apoptotic chondrocytes was significantly lower in the experimental group 1 at 3 weeks after transplantation (P<0.05). These results indicate that autologous oxygen-delivering biomimetic nanoscaffold composited with chondrocytes to repair different types of temporomandibular joint condylar fractures presents some different outcomes.
8.Effect of Comprehensive Rehabilitation on Shoulder Subluxation of Stroke Patients with Hemiplegia
Ai-dong LI ; Hong-tao LIU ; Zong-qing HUANG ; Yingtai DENG ; Guifang WAN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(8):673-674
ObjectiveTo observe the effect of comprehensive rehabilitation on shoulder subluxation of patients with stroke hemiplegia.MethodsComprehensive rehabilitation was practiced on 20 hemiplegic post-stroke patients with shoulder subluxation, including putting injured limb at correct position, keeping the range of passive motion, resistance exercise of scapula muscle group, and shoulder sting. The therapeutic effect was evaluated after 6 weeks treatment.ResultsThere was a significant difference in the shoulder subluxation and movement function of the upper extremities between pre-therapy and after therapy ( P<0.05~0.01).ConclusionComprehensive rehabilitation has obvious therapeutic effect on shoulder subluxation and motor dysfunction of patients with stroke hemiplegia.
9.Rosuvastatin attenuates mucus secretion in a murine model of chronic asthma by inhibiting the gamma-aminobutyric acid type A receptor.
Tao ZHU ; Wei ZHANG ; Dao-xin WANG ; Ni-wen HUANG ; Hong BO ; Wang DENG ; Jia DENG
Chinese Medical Journal 2012;125(8):1457-1464
BACKGROUNDAsthma is a chronic inflammatory disease characterized by reversible bronchial constriction, pulmonary inflammation and airway remodeling. Current standard therapies for asthma provide symptomatic control, but fail to target the underlying disease pathology. Furthermore, no therapeutic agent is effective in preventing airway remodeling. A substantial amount of evidence suggests that statins have anti-inflammatory properties and immunomodulatory activity. In this study, we investigated the effect of rosuvastatin on airway inflammation and its inhibitory mechanism in mucus hypersecretion in a murine model of chronic asthma.
METHODSBALB/c mice were sensitized and challenged by ovalbumin to induce asthma. The recruitment of inflammatory cells into bronchoalveolar lavage fluid (BALF) and the lung tissues were measured by Diff-Quik staining and hematoxylin and eosin (H&E) staining. ELISA was used for measuring the levels of IL-4, IL-5, IL-13 and TNF-α in BALF. Periodic acid-Schiff (PAS) staining was used for mucus secretion. Gamma-aminobutyric acid type A receptor (GABAAR) β2 expression was measured by means of immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.
RESULTSRosuvastatin reduced the number of total inflammatory cells, lymphocytes, macrophages, neutrophils, and eosinophils recruited into BALF, the levels of IL-4, IL-5, IL-13 and TNF-α in BALF, along with the histological mucus index (HMI) and GABAAR β2 expression. Changes occurred in a dose-dependent manner.
CONCLUSIONSBased on its ability to reduce the inflammatory response and mucus hypersecretion by regulating GABAAR activity in a murine model of chronic asthma, rosuvastatin may be a useful therapeutic agent for treatment of asthma.
Animals ; Asthma ; drug therapy ; metabolism ; Chronic Disease ; Disease Models, Animal ; Female ; Fluorobenzenes ; pharmacology ; therapeutic use ; GABA-A Receptor Antagonists ; pharmacology ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Lung ; chemistry ; Mice ; Mice, Inbred BALB C ; Mucus ; secretion ; Pyrimidines ; pharmacology ; therapeutic use ; Receptors, GABA-A ; analysis ; Rosuvastatin Calcium ; Sulfonamides ; pharmacology ; therapeutic use
10.The comparative study of cognitive functions between familial and sporadic patients with first-episode schizophrenia
Mei WANG ; Hong DENG ; Wei DENG ; Tao LI ; Mingli LI ; Na LI ; Yin LIN ; Hongyan REN ; Zhaohua HUANG ; Yinfei LI ; Wei LEI ; Yuanyuan HAN
Chinese Journal of Nervous and Mental Diseases 2014;(4):218-222
Objective To investigate the difference in cognitive functions between first-episode schizophrenia pa-tients with and without family history. Methods One hundred twenty-seven patients with first-episode schizophrenia were recruited, including 40 patients with family history and 87 sporadic patients. Ninety-six matched normal subjects served as controls. Seven subscales of the Wechsler Adult Intelligence Scale-Revised in China (WAIS-RC) were used to assess the cognitive functions of all subjects. Positive and Negative Symptom Scale (PANSS) was used to assess patients ’ symptoms. The relationship between clinical symptoms and cognitive deficits was analyzed. Results There was no signifi-cant difference in the PANSS scores between familial patients and sporadic patients [(91.51±14.07) vs. (87.23±16.37), P>0.05]. The scores of full intelligence quotient (IQ), verbal IQ and operation IQ were lower in patient groups than in con-trol group (all P<0.05). The score of operation IQ was lower in sporadic patients than in familial patients (P<0.05). The scores of PANSS (r=-0.43, P=0.01), positive symptoms (r=-0.32, P=0.04) and negative symptoms (r=-0.38, P=0.02) were negatively correlated with operation IQ, and the scores of PANSS(r=-0.41,P=0.01) and positive symptoms(r=-0.54, P<0.01) were negatively correlated with block rectification scores in sporadic patient group but not in the familial patient group (all P>0.05). Conclusion The patients with schizophrenia have significant intelligence deficits in the early stages. Cognitive deficits are associated with the disease severity in sporadic patients while are independent of clinical symptoms in familial patients.