1.Analysis of microbial diversity in the midgut of Haemaphysalis longicornis infected with severe fever with thrombocytopenia syndrome virus
CHEN Chen ; YUAN Chuan-fei ; KANG Le ; XIA Qian-feng
China Tropical Medicine 2023;23(4):347-
Abstract: Objective To investigate the composition and diversity of midgut microbial community of Haemaphysalis longicornis infected with severe fever with thrombocytopenia syndrome virus (SFTSV). Methods The midgut DNA of three group Haemaphysalis longicornis infected with SFTSV was extracted, and the 16S rDNA gene of the sample was sequenced by HiSeq platform. The composition and diversity of endosymbiotic microbial community were clarified by OTU cluster analysis and alpha diversity analysis. Results The midgut microbial clusters of the three groups infected with SFTSV were 143, 113, 163 OTUs respectively; the sparsity curve and abundance grade curve showed that the data had sufficient sequencing depth, and the midgut of Haemaphysalis longicornis infected with SFTSV was rich in microbial composition, but the species distribution was uneven. The analysis of microbial community composition showed that Proteobacteria, Firmicutes and Actinobacteria were the main dominant bacteria at the phyla level. At the class level, Gammaproteobacteria, Bacilli, Betaproteobacteria and Actinomycetia were the main dominant bacteria. At the order level, Legionellales, Bacillales, Burkholderiales and Actinomycetales were the main dominant orders. At the family level, Coxiellaceae, Bacillaceae, Moraxellaceae and Rhodococcaceae were the main dominant families. At the genus level, the relative abundance of Coxiella was the highest, followed by Aeribaillus and Azonexus. Alpha diversity analysis showed that the average Shannon index was 139.67, the average Simpson index was 0.48, the average Chao index was 145.06, and the average ACE index was 147.11. Conclusions The species diversity of intestinal microorganisms in Haemaphysalis longicornis infected with SFTSV is rich. The results provide a basis for further exploring the interaction between intestinal microbes of Haemaphysalis longicornis and SFTSV and developing new ideas for the prevention and control of ticks and tick-borne diseases.
2.Determination of prulifloxacin active metabolite in human plasma and urine by RP-HPLC
Juan HE ; Yong-Chuan CHEN ; Qing DAI ; Pei-Yuan XIA ;
Chinese Journal of Infection and Chemotherapy 2007;0(01):-
Objective To establish a RP-HPLC method for determining the concentration of prulifloxacin active metabolite in human plasma and urine.Methods The supernatant obtained by centrifugation after the sample was precipitated with methanol- acetonitrile (1:1) was chromatographically separated on a Diamonsil C_(18)(250 mm?4.6 mm,5?m) using a mobile phase con- sisting of acetonitrile and 0.05 mol/L potassium dihydrogen phosphate (pH2.2) containing 1% tetrabutylammonium bromide. The solutions of 20:80 (V/V) and 12:88 (V/V) at a flow rate of 1.0 mL/min and 1.6 mL/min were used for plasma and u- rine, respectively.Then the samples were assayed at wavelength of Ex 280 nm and Em 425 nm.Results The linear range for prulifloxacin active metabolite in plasma and urine were 0.005-5 mg/L (r=0.9999) and 0.05-5 mg/L(r=0.9999)with a low- er limit of quantitation of 0.002 mg/L and 0.01 rag/L, respectively.In plasma, the relative recovery ranged from 100.64% to 101.00% at the concentration of 5.00, 0.50 and 0.05 mg/L and within-day and between-day precisions were less than 2.5% and 4.6% respectively.Meanwhile, the relative recovery ranged from 97.20% to 100.20% at the concentration of 2.50, 0.50 and 0.10 mg/L in urine.The within-day and between-day precisions were lower than 1.3% and 4.3%, respectively.The method had been successfully used for the pharmacokinetic studies of a prulifloxacin formulation after oral administration to healthy volunteers.Conclusions The present method is simple, rapid, accurate, reproducible and suitable for the pharmacoki- netic study of prulifloxacin in humans.
3.Study of Apoptosis and Gene Expression Pattern Related to Human Degenerative Intervertebral Disc Tissue
Ming HU ; Yuan-zheng MA ; Chuan-sen ZHANG ; Daoyuan CHEN ; Xing CHEN ; Xiaojun CAI ; Yibing BAI
Chinese Journal of Rehabilitation Theory and Practice 2006;12(6):484-486
ObjectiveTo investigate the relationship between the degenerative mechanisms of lumbar intervertebral disc (LID) and apoptosis.MethodsThe total RNAs were isolated from human LID tissues. Both the mRNAs from the degeneration and normal LID were reversely transcribed to the cDNAs. The cDNAs were labeled with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After high-stringent washing, the cDNA microarray was scanned for the fluorescent signals and analyzed by computer image analysis. The apoptotic status and the expression of Bcl-2 and Bax in 12 cases of degenerative LID and 10 cases of normal LID were detected with TdT-mediated dUTP-biotin nick end labeling (TUNEL) and immunohistochemistry methods.ResultsAmong the 4096 targets, there were 10 genes related to apoptosis. The expression related to Bax protein gene was up-regulated and it was down-regulated for Bcl-2 protein. In group of normal LID, the average apoptotic index (AI) was (24.897±3.620); percentage of Bcl-2 positive cells was (31.440±4.150)%; percentage of Bax positive cells was (29.372±2.588)%, average optical density (OD) values of positive particles were (0.183± 0.010 ), ( 0.203 ±0.012) and (0.169±0.005) respectively. In group of degenerative LID, the average AI was (49.232±3.440); percentage of Bcl-2 positive cells was (18.239±2.470)%; percentage of Bax positive cells was (52.349±3.764)%; average OD values of positive particles were (0.152±0.003), (0.310±0.008) and (0.262±0.014) respectively. There were significantly differences in AI and expressions of Bcl-2 and Bax proteins between normal LID and degenerative LID (P<0.05).ConclusionCell apoptosis plays an important role in the process of LID degeneration. Both Bcl-2 and Bax take part in the occurrence and progression of LID.
4.Inflammation is involved in the organ damage induced by sinoaortic denervation in rats
Chuan ZHANG ; Hong CHEN ; Hehui XIE ; He SHU ; Wenjun YUAN ; Dingfeng SU
Academic Journal of Second Military Medical University 2004;25(4):388-388
Objective:The present study was designed to test the hypothesis that inflammation is involved in the end-organ damage(EOD) induced by sinoaortic denervation(SAD) in rats.Method:SAD was performed in male Sprague-Dawley rats at the age of 10 weeks.Under anaesthesia,aortic nerves were cut and the sinus region of the carotid artery was stripped and painted with 10% phenol.Pathological evaluation of EOD and the determination of plasma or tissue levels of the factors related to inflammation,including thromboxane B2(TXB2) interleukin-1(IL-1),tumour necrosis factor α(TNF-α) and reactive oxygen species(ROS) were performed at 16 weeks after SAD.Pathological evaluation of EOD included heart weigh ratio,myocardial and blood vessel hydroxyproline and collagen volume fraction,glomerular injury score and number of infiltrating inflammatory cells.Indomethacin(20 mg/kg per day,orally) or vitamin E(100 mg/kg per day,orally) was administered for 12 weeks,beginning from4 weeks after SAD,to observe their effects on SAD-induced EOD.Results:There were significant fibrosis and inflammatory infiltration in the myocardium and blood vessels,represented by higher hydroxyproline and collagen volume fraction,and a large amount of inflammatory cells in the tissues of SAD rats.Heart weight and kidney glomerular injury score were significantly higher in ed significantly after SAD.Indomethacin and vitamin E significantly decreased the contents of some factors related to inflammation in SAD rats.Both drugs also alleviated myocardial and vessel fibrosis,inflammatory infiltration and kidney damage.Conclusion:Inflammation is involved in the organ damage induced by SAD in rats.
5.Effect of shenhuang ointment in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of Qi stagnation blood stasis syndrome: a clinical observation.
Xing WEI ; Hua-seng QIU ; Qi ZHANG ; De-chuan LI ; Yuan-shui SUN ; Gang LI ; De-xing CHEN ; Bin ZHANG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(6):661-665
OBJECTIVETo observe the effect and safety of plastering Chinese Compound Shenhuang Ointment (CSO) at Shenque (RN8) in promoting the rehabilitation of postoperative gastrointestinal dysfunction patients of qi stagnation blood stasis syndrome (QSBSS).
METHODSA prospective, multi-centered, randomized, double-blinded, controlled trial was conducted in 220 postoperative gastrointestinal dysfunction patients of QSBSS. They were randomly assigned to two groups, the CSO group (110 cases) and the placebo group (110 cases). CSO was plastered at Shenque (RN8) for 5 days after operation. The time of exhaustion, defecation, the recovery of intestinal peristalsis, integrals of TCM syndrome, and serum levels of motilin (MOT)and somatostatin (SS) were observed.
RESULTSCompared with the placebo group, the condition of exhaustion and defecation, the recovery of intestinal peristalsis on the 3rd day after operation was all improved (P < 0.05). The integrals of TCM syndrome at day 2, 3, and 4 were more significantly lowered in the CSO group than in the placebo group (P < 0.01, P < 0.05). The total effective rate of TCM syndrome was 95.3% in the CSO group, better than that in the placebo group (91.8%, P < 0.05). Compared with the placebo group, the serum MOT level increased and the serum SS level decreased at day 5 after operation in the CSO group (P < 0.05).
CONCLUSIONSThe plastering of CSO at Shenque (RN8) could advance the time of exhaustion and defecation, and improve patients' clinical symptoms. And patients could tolerate well.
Aged ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastrointestinal Diseases ; drug therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ointments ; Postoperative Period ; Prospective Studies
6.An invitro biomechanical study of treatment for lumbar spondylolithesis using nail-grooved tail steel plate and intervertebral implant (Wendeng Fusion Cage, WDFC).
Yuan-chao TAN ; Chuan-guang JU ; Xiu-chen SUN ; Zeng-mei WANG
China Journal of Orthopaedics and Traumatology 2008;21(8):566-569
OBJECTIVETo evaluate the biomechanics of treatment for lumbar spondlolisthesis using nail-grooved tail steel plate and WDFC (Wendeng Fusion Cage) implant.
METHODSThere were nine permanent waist-sacrum wet bone (L3-S3) in 1 to 2 clay-cold hours including 6 men and 3 women. They were seldom separated into 3 groups, which were fixed by nail-grooved fail plus WDFC. The model was separate into two kinds for single and across. With electrometry, deal experiment date with Graftool software. Each piece should be tested twice respectively.
RESULTSThe single and across segment non-destructive compression experiment. No-mid-compression from 0 to 750 N,the related coefficient and curves had no obvious change on inclined rate. In the single segment curvedly serial experiment, the stress at all point measured by two sides steel plate-was mostly linear growth. In the across segment curvedly serial experiment, the inclined rate become big and appear anisomerous.
CONCLUSIONIt's proved by biomechanics that the steel plates with single furrow and cylinder wing plus WDFC has a good stability to cure lumbar vertebra slips.
Adult ; Biomechanical Phenomena ; Bone Nails ; Bone Plates ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Spinal Fusion ; instrumentation ; Spondylolisthesis ; physiopathology ; surgery
7.Effects of extremely low frequency magnetic fields on hydrolysis of F0F1-ATPases and their relationship with turnover rates of F1.
Chuan-Fang CHEN ; Yuan-Bo CUI ; Jia-Chang YUE
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(6):327-331
OBJECTIVETo study the effects of extremely low frequency sinusoidal magnetic fields on hydrolysis of F(0)F(1)-ATPase and its mechanism.
METHODSThe F(0)F(1)-ATPases which was localized on the outer surface of chromatophores were prepared from the cells of Rhodospirillum rubrum and were exposed to 0.1 approximately 0.5 mT, 4.7 approximately 96.0 Hz magnetic fields.
RESULTSThe hydrolysis activity of F(0)F(1)-ATPase was stimulated by 0.5 mT, 4.7, 12.0, 60.0, 72.0, 84.0 and 96.0 Hz magnetic fields respectively and inhibited by 0.5 mT, 24.0 Hz magnetic field (P < 0.05); 0.3 mT, 4.7, 24.0 and 60.0 Hz magnetic fields also distinctly affected F(0)F(1)-ATPases activity respectively (P < 0.05), whereas 0.1 mT exposure caused no significant changes on that activity. When the hydrolysis activity of the F(0)F(1)-ATPases was inactivated by its inhibitor DCCD, the 0.5 mT, 24.0 Hz magnetic field still inhibited the hydrolysis activity of the F(0)F(1)-ATPase and 0.5 mT, 60.0 Hz magnetic field also had stimulating effects (P < 0.05).
CONCLUSIONThe effects of magnetic fields on the hydrolysis activity of the F(0)F(1)-ATPases depend on not only magnetic frequency but also magnetic intensity. The threshold of magnetic intensity is between 0.1 mT and 0.3 mT. F(0)F(1)-ATPases, especially F1-portion may be an end-point of magnetic fields.
Hydrolysis ; radiation effects ; Magnetic Fields ; adverse effects ; Proton-Translocating ATPases ; metabolism ; Rhodospirillum rubrum ; enzymology
8.Brain CT texture classification with tree-structured wavelet transform.
Di CHEN ; Ping ZHOU ; Chuan-fu LI ; Kang-yuan ZHOU
Chinese Journal of Medical Instrumentation 2007;31(4):239-241
This paper proposes a tree-structured wavelet algorithm for brain CT texture feature extraction. This algorithm is used to analyse subimages automatically and to choose the optimum subimage for feature extraction. Experimental results show that this algorithm can improve the performance of brain CT texture feature extraction.
Algorithms
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Brain
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diagnostic imaging
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Humans
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Image Processing, Computer-Assisted
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Tomography, X-Ray Computed
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methods
9.Prokaryotic Expression and Immunological Reactivity of Rotavirus NSP6
Chuan-Yin LI ; Yao-Chun FAN ; Yu-Ling WEN ; Yan ZHANG ; Hai-Tao WEI ; Yuan-Ding CHEN ;
China Biotechnology 2006;0(09):-
Little is known about the non-structural protein 6(NSP6)of rotavirus.This report describes expression of the NSP6 of a group A human rotavirus strain TB-Chen in bacteria,and its immunological properties and cellular distribution.The results showed that the recombinant NSP6(rNSP6)was expressed in high efficiency without any other proteins fused(possesses about 34.2% of total bacterial proteins).rNSP6 elicited mono-specific antibodies in immunized guinea pigs and the antibodies could react with the rNSP6 itself and the viral NSP6 proteins synthesized in SA11-or Wa-infected MA104 cells in Western blot and immunofluorescence assay.The NSP6 distributed evenly in the cytoplasm mainly around the nucleus of virus-infected cells,no viroplasm-like gathering observed;The top amount of NSP6 synthesized in SA11-infected cells or Wa-infected cells could be detected at 12h after infection.This is the first report about the high expression of entire NSP6(without any other proteins fused)in prokaryotic expression system and detection of NSP6 synthesis in virus infected cells by immunofluorescence assay.The results are important to understand the structure,biological properties and further application of the NSP6.
10.Inflammation is involved in the organ damage induced by sinoaortic denervation in rats
Chuan ZHANG ; Hong CHEN ; He-Hui XIE ; He SHU ; Wen-Jun YUAN ; Ding-Feng SU
Academic Journal of Second Military Medical University 2004;25(4):388-388
Objective:The present study was designed to test the hypothesis that inflammation is involved in the end-organ damage(EOD) induced by sinoaortic denervation(SAD) in rats.Method:SAD was performed in male Sprague-Dawley rats at the age of 10 weeks.Under anaesthesia,aortic nerves were cut and the sinus region of the carotid artery was stripped and painted with 10% phenol.Pathological evaluation of EOD and the determination of plasma or tissue levels of the factors related to inflammation,including thromboxane B2(TXB2) interleukin-1(IL-1),tumour necrosis factor α(TNF-α) and reactive oxygen species(ROS) were performed at 16 weeks after SAD.Pathological evaluation of EOD included heart weigh ratio,myocardial and blood vessel hydroxyproline and collagen volume fraction,glomerular injury score and number of infiltrating inflammatory cells.Indomethacin(20 mg/kg per day,orally) or vitamin E(100 mg/kg per day,orally) was administered for 12 weeks,beginning from4 weeks after SAD,to observe their effects on SAD-induced EOD.Results:There were significant fibrosis and inflammatory infiltration in the myocardium and blood vessels,represented by higher hydroxyproline and collagen volume fraction,and a large amount of inflammatory cells in the tissues of SAD rats.Heart weight and kidney glomerular injury score were significantly higher in ed significantly after SAD.Indomethacin and vitamin E significantly decreased the contents of some factors related to inflammation in SAD rats.Both drugs also alleviated myocardial and vessel fibrosis,inflammatory infiltration and kidney damage.Conclusion:Inflammation is involved in the organ damage induced by SAD in rats.