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ObjectiveTo evaluate the clinical efficacy of TDP （specific electromagnetic wave） combined with Osteoking in the treatment of knee osteoarthritis of Qi stagnation and blood stasis type. MethodA total of 104 patients with knee osteoarthritis of Qi stagnation and blood stasis type， who received conservative therapy in The Third People's Hospital of Xinjiang Uygur Autonomous Region from July 2019 to December 2021， were randomized into the control group and study group with the random number table method， 52 cases in either group. The control group was treated with TDP， and the study group with TDP and Osteoking. The treatment lasted 1 week for both groups， with 1-month follow-up. Subjective indexes of visual analog scale （VAS） score and Western Ontario McMaster Universities Osteoarthritis Index （WOMAC） score， and objective indexes of visual tenderness index and visual knee range of motion were determined before and after treatment to evaluate the pain and functions of patients. Traditional Chinese medicine （TCM） syndrome score was calculated. The serum erythrocyte sedimentation rate （ESR） and high sensitivity C-reactive protein （hs-CRP） were detected before and after treatment， and the total clinical effective rate was calculated. ResultBefore treatment， the baseline information and all the scores of the two groups were comparable. After treatment， the VAS score， WOMAC score， tenderness index， knee range of motion， and TCM syndrome score were improved in both groups （P<0.01）. After the treatment， the VAS score and WOMAC score of the study group were lower than those of the control group （P<0.01） and the improvement of tenderness index in the study group was better than that in the control group （P<0.05）. The knee range of motion in the study group was better than that in the control group （P=0.061）. The TCM syndrome score of study group was lower than that of control group （P<0.01） after treatment. The post-treatment serum ESR and hs-CRP level in the two groups decreased significantly after treatment， and the study group were lower than those of the control group （P<0.01）. The total clinical effective rate of the study group was 90.4%（47/52）， as compared with the 53.8%（28/52） in the control group （P<0.05）. No obvious adverse events occurred during treatment in both groups. ConclusionThe clinical efficacy of TDP combined with Osteoking in the treatment of knee osteoarthritis of Qi stagnation and blood stasis type is remarkable， which can improve knee pain and functions， alleviate TCM syndrome， and reduce inflammatory indexes， with high safety.
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Objective:To research the metabolomic alterations of human lung bronchial epithelial cells infected with human rhinovirus 1B (HRV1B).Methods:Untargeted metabolomics was used to determine the metabolomic alterations in human lung bronchial epithelial cells (BEAS-2B) 6 h, 12 h and during the dynamic process (6 h∶12 h) after HRV1B infection.Results:A total of 93 differentially significant metabolites (DSMs) (47 DSMs were up-regulated and 46 DSMs were down-regulated) and 88 DSMs (37 DSMs were up-regulated and 51 DSMs were down-regulated) at post infection of HRV1B in BEAS-2B at 6 h or 12 h, respectively. A total of 30 DSMs (12 DSMs were up-regulated and 18 DSMs were down-regulated) in a dynamic process (6 h∶12 h) after HRV1B infection. Unknown metabolites took up most proportions. The trends of fatty acid, lipid, amino acid, nucleotide and carbohydrate were increased along with the prolonging of HRV1B infection. DSMs such as Diisononyl phthalate was co-detected DSMs among three groups.Conclusions:Metabolites such as fatty acid, lipid, amino acid, nucleotide and carbohydrate of BEAS-2B cells are changed induced by HRV1B infection.
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Objective:To explore the application and value of metagenomic next-generation sequencing (mNGS) in refractory infection after organ transplantation.Methods:A case report discussed about a patient with lumbar spine infection after kidney transplantation and the relevant literature was reviewed. The recipient was a 63-year-old man with low back pain after kidney transplantation. Lumbar spine magnetic resonance imaging showed lumbar spine infection. Multiple operations plus antibacterial and antituberculosis treatments were ineffective. Before and after treatment, numerous tests of traditional pathogenic microorganisms failed to detect any positive bacteria.Results:The detection of lumbar secretion by mNGS suggested aspergillus infection. The symptoms improved after dosing of voriconazole.Conclusions:The incidence of fungal infection of lumbar spine is low. The imaging manifestations are non-typical so that it is easy to misdiagnose. mNGS helps to timely diagnose and guide treatment. With a review of the literature, mNGS has some application value for some difficult and rare infectious diseases.
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OBJECTIVE：To establish the fingerprint of crude/vinegar-processed Corydalis yanhusuo decoction pieces and their dispensing granules，and to determine the contents of five alkaloids （protopine，tetrahydropalmatine，corydaline，berberine hydrochloride，palmatine hydrochloride ）. METHODS ：HPLC method was adopted. The determination was performed on Agilent TC-C18 column with mobile phase consisted of 0.1％ phosphoric acid solution-methanol （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm，and the column temperature was 30 ℃. The sample size was 10 μL. Using palmatine hydrochloride as reference ， Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition） was used to establish the fingerprint of 11 059） batches of C. yanhusuo decoction pieces ，7 batches of crude . yanhusuo dispensing granules ， 12 batches of vinegar- processed C. yanhusuo decoction pieces and 11 batches of vinegar-processed C. yanhusuo dispensing granules. The same HPLC method was adopted to determine the contents of protopine， tetrahydropalmatine， corydaline， berberine hydrochloride and palmatine hydrochloride in 41 batches of crude/ vinegar-processed C. yanhusuo decoction pieces and their dispensing granules. RESULTS ：There were 12 and 20 common peaks for crude C. yanhusuo decoction pieces and its dispensing granules ，and 14 and 16 common peaks for vinegar-processed C. yanhusuo decoction pieces and its dispensing granules. The similarity of each batch of same type were 0.529-0.981，0.342-0.985， 0.711-0.999，0.437-0.998，respectively. The linear range of protopine ，tetrahydropalmatine，corydaline，berberine hydrochloride and palmatine hydrochloride were 1.9-38.0，2.0-40.0，2.2-44.0，2.6-52.0，2.3-46.0 μg/mL（R2＞0.999 0）. The recoveries were 100.12％-100.98％（RSD＝1.05％-1.90％，n＝9）. RSDs of precision ，reproducibility，stability（24 h）and durability tests were all lower than 2.0％. The average contents of five alkaloids in different batches of crude/vinegar-processed C. yanhusuo decoction pieces and its dispensing granules were 0.24-0.46，0.37-0.82，0.24-0.58，0.07-0.75，0.24-0.76 mg/g. RSDs were 12.27％-147.48％. CONCLUSIONS：The fingerprint of crude/vinegar-processed C. yanhusuo decoction pieces and its dispensing granules is established successfully. The similarities of fingerprint are different before and after processing with vinegar ，and the contents of five alkaloids in C. yanhusuo decoction pieces and its dispensing granules are greatly different.
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Objective@#Remuzzi scoring system is utilized for assessing the degree of renal tissue damage in donors with hypertensive cerebral hemorrhage and donors with brain trauma after cardiac death. To explore the prognosis of hypertensive cerebral hemorrhage donor kidney in renal transplant recipients.@*Methods@#The kidney donated by DCD between January 1, 2016 to June 1, 2018 were retrospectively reviewed. Pathological biopsy was performed before transplantation and hematoxylin-eosin (HE) staining after sectioning. The degree of renal tissue lesions was evaluated by Remuzzi scoring system. According to the source of donor kidney, they were divided into two groups of donors with heart failure due to hypertensive cerebral hemorrhage (HCH) and those with brain trauma (BT). Both groups of donor kidneys were preserved by low-temperature machine perfusion. The immunosuppressive regimen was identical in both groups. The prognosis of two groups was compared by serum creatinine (Scr) at Month 1/6/12 post-operation and cumulative graft survival rate over a follow-up period of 12-36 months.@*Results@#The renal Remuzzi score of HCH donors was significantly higher than that of BT donors. The maximal creatinine clearance rate was significantly lower than that of BT donors [(86.8±27.8) vs (115.4±23.2) ml/min, P<0.05]. At 1/6/12 months post-transplantation, serum creatinine levels were (76.1±18.5), (72.4±16.2) and (71.4±16.8) μmol/L in BT group and (160.3±33.4), (154.3±32.6) and (146.4±29.1) μmol/L in HCH group. The SCr in BT group at 1/6/12 months was lower than that in HCH group (P<0.05). Kaplan-meier analysis showed no significant inter-group difference in graft survival between two groups over a follow-up period of 12 to 36 months (Log-Rank test, P=0.485).@*Conclusions@#No significant difference exists in short-term survival rate of kidneys from HCH and BT donors. The recipients of HCH donor's kidney have higher serum creatinine levels than those of BT donors. Selective use of kidney transplants in patients with cardiac death caused by HCH may greatly reduce the waste of donor kidney and improve the quality-of-life of patients with end-stage renal disease.
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Objective Remuzzi scoring system is utilized for assessing the degree of renal tissue damage in donors with hypertensive cerebral hemorrhage and donors with brain trauma after cardiac death .To explore the prognosis of hypertensive cerebral hemorrhage donor kidney in renal transplant recipients .Methods The kidney donated by DCD between January 1 ,2016 to June 1 ,2018 were retrospectively reviewed .Pathological biopsy was performed before transplantation and hematoxylineosin (HE) staining after sectioning . The degree of renal tissue lesions was evaluated by Remuzzi scoring system .According to the source of donor kidney ,they were divided into two groups of donors with heart failure due to hypertensive cerebral hemorrhage (HCH) and those with brain trauma (BT) . Both groups of donor kidneys were preserved by low-temperature machine perfusion . The immunosuppressive regimen was identical in both groups .The prognosis of two groups was compared by serum creatinine (Scr) at Month 1 /6/12 post-operation and cumulative graft survival rate over a follow-up period of 12-36 months .Results The renal Remuzzi score of HCH donors was significantly higher than that of BT donors .The maximal creatinine clearance rate was significantly lower than that of BT donors [(86 .8 ± 27 .8 ) vs (115 .4 ± 23 .2 ) ml/min , P < 0 .05 ] . At 1 /6 /12 months posttransplantation ,serum creatinine levels were (76 .1 ± 18 .5 ) , (72 .4 ± 16 .2 ) and (71 .4 ± 16 .8) μmol/L in BT group and (160 .3 ± 33 .4) ,(154 .3 ± 32 .6) and (146 .4 ± 29 .1) μmol/L in HCH group .The SCr in BT group at 1 /6 /12 months was lower than that in HCH group ( P < 0 .05 ) . Kaplan-meier analysis showed no significant inter-group difference in graft survival between two groups over a follow-up period of 12 to 36 months (Log-Rank test , P = 0 .485 ) .Conclusions No significant difference exists in short-term survival rate of kidneys from HCH and BT donors . The recipients of HCH donor's kidney have higher serum creatinine levels than those of BT donors . Selective use of kidney transplants in patients with cardiac death caused by HCH may greatly reduce the waste of donor kidney and improve the quality-of-life of patients with end-stage renal disease .
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Many malignant tumors, most commonly small cell lung cancer, merge with syndrome of inappropriate secretion of antidi-uretic hormone (SIADH). SIADH is caused by the neuroendocrine function of tumor cells or is a side effect of chemotherapeutics. As a chemotherapeutic side effect, SIADH presents as hyponatremia. Therapy for SIADH includes both treatment for primary diseases and hyponatremia. Given that SIADH-induced hyponatremia aggravates the prognosis and shortens the survival time of patients with malig-nant tumors, SIADH is an independent risk factor for prognosis. Therefore, the early diagnosis, monitoring, and treatment of SIADH will improve the prognosis of patients with malignant tumors.
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Objective To observe the changes of serum heart-type fatty acid-binding protein (h-FABP) and brain natriuretic peptide (BNP) for assessing the clinical values in children with chronic heart failure (CHF).Methods A total of 50 patients with CHF,including 22 endocardial fibroelastosis (EFE) cases and 28 dilated cardiomyopathy (DCM) cases,were enrolled.Thirty healthy children were enrolled as controls.The concentrations of serum h-FABP and BNP were measured by enzyme-linked immunosorbent assay (ELISA),and the left ventricular end-systolic diameter (LVESD),left ventricular end-diastolic diameter (LVEDD),left ventricular ejection fraction (LVEF),left ventricular fractional shortening (LVFS),and cardiac index (CI) were measured by echocardiography.Results The concentrations of serum h-FABP and BNP in patients with CHF were significantly higher than those in control group (22.68 ±4.18 ng/mL vs.6.31 ± 1.72 ng/mL;565.52±179.64 pg/mLvs.31.22±9.83 pg/mL,all,P ＜ 0.01),and h-FABP and BNP were positively correlated with the degree of heart failure (all,P ＜0.01),and they were both higher in group EFE and DCM than those in control group (all,P＜0.01),but there was no statistically significant difference between the two groups (P ＞ 0.05).The h-FABP concentration in CHF patients was positively correlated with BNP ((r =0.782,P ＜0.01).The h-FABP concentration in CHF patients was positively correlated with LVESD,LVEDD (r =0.626,O.662,respectively;all,P＜0.01),and negatively correlated with LVEF,LVFS,and CI (r =-0.652,-0.636,-0.709,respectively;all,P ＜ 0.01).The BNP concentration was also positively correlated with LVESD,LVEDD (r =0.733,0.689,respectively;all,P ＜0.01),and negatively correlated with LVEF,LVFS,and CI (r=-0.745,-0.612,-0.791,respectively;all,P＜0.01).Conclusions Serum concentrations of h-FABP and BNP can be used as biomarkers to evaluate the severity of chronic heart failure.
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Objective To make students at different levels not only learn theoretical knowledge of acupuncture-moxibustion manipulation skills but also achieve full operating training, strengthen the combination of theory and practice and improve the teaching quality of clinical disciplines by applyingmultimedia-subject participationteaching method to acupuncture-moxibustion manipulation techniques in the experimental teaching of acupuncture and moxibustion. Method The students were randomized, according to grade, into experiment (n=114) and control (n=115) groups. The teaching content was the same in the two groups. The experiment group received multimedia-subject participationteaching method. After the multimedia teaching, the students did an independent demonstration and made a self-evaluation, the teachers made a comment and the students did independent practice again in groups. The control group received conventional theory teaching and did practice under demonstration in the experimental lesson. The examination and questionnaire were used as assessment indicators to evaluate the effect ofmultimedia-subject participationteaching method applied to acupuncture-moxibustion technique teaching. Result Statistical analysis was made using the SPSS11.5 statistical software package. The independent samples t-test is used for a between-groups comparison. The statistical result showed that satisfaction was 100%in the experiment group and 75.7%in the control group;there was a statistically significant difference between the two groups (P<0.05). Conclusion Multimedia-subject participationteaching method is an innovation in acupuncture-moxibustion technique teaching and helps the students to better master acupuncture-moxibustion manipulation skills. Mastery of theoretical knowledge before real practice is the key to proficient manipulation. The combination of the two in classroom teaching yields twice the result with half the effort. Carrying out multimedia-subject participation teaching method will help to further deepen the teaching reform and to improve the level and quality of“Acupuncture and Moxibustion”teaching.
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Aim To investigate the impact of CYP3 A5 genetic polymorphism on modified releasing tacrolimus pharmacokinetics in Chinese stable renal transplant re-cipients. Methods Pharmacokinetics of once daily-ta-crolimus( tac-q. d. ) and twice daily-tacrolimus( tac-b. i. d. ) were determined by CLIA, CYP3A5 genotype was measured by PCR-RFLP. Each 10 patients receiv-ing tac-q. d. and tac-b. i. d. respectively were en-rolled, and each 5 patients receiving tac-q. d. were matched to poor metabolizer ( PM ) and extensive me-tabolizer ( EM ) group respectively according to CYP3A5 genotypes. Results AUC0~24 h for tac-q. d. was 1. 78 folds higher than AUC0~12 h for tac-b. i. d. , and dose-adjusted C0 was 40% lower for tac-q. d. than for tac-b. i. d. There were no significant differences for other parameters between the two groups; Cmax, AUC0~24 h and C0 were 1. 75, 1. 96 and 2. 49 folds higher for PM than for EM, and dose-adjusted Cmax, AUC0~24 h and C0 were 1. 80, 2. 34 and 2. 64 folds higher for PM than for EM. There were good correla-tions between AUC0~24 h and C0 for tac-q. d. Conclu-sion Conversion from tac-b. i. d. to tac-q. d. results in requirement of increased tacrolimus dose and detec-tion of CYP3A5 genotype, which is necessary for ensu-ring C0 in the range of therapeutic window.
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This study was aimed to summarize the adverse drug reaction (ADR) caused by the toxicity of Tripterygium Wilfordiiand its preparations, in order to explore possible relationship betweenTripterygium Wilfordiifactors and reported ADR. Relevant articles on toxicity ofTripterygium Wilfordiiand its preparations were systematically searched in 5 databases, including the Pubmed, CNKI,Wanfang Data, VIP Data and Sinomed from the database was established until Feb 25th, 2014. And then, the randomized controlled trials (RCTs) were systematically collected, analyzed and summarized. The results showed that there were 260 RCTs with 13301 patients included. The outcome of data analysis showed that ADR rates of digestive system and reproductive system of RCT1 and RCT2 were different. ADR rates (per hundred people) in RCT1 and RCT2 were as follows: digestive system were 14.73 and 12.26, reproductive system were 8.25 and 8.00, liver was 6.50 and 5.66, kidneys were 6.79 and 3.03, blood system were 6.73 and 6.50, cardiovascular system were 2.35 and 0.67, skin and mucous system were 11.42 and 4.78, respectively. Articles on rheumatoid arthritis (RA) of both RCT1 and RCT2 were the highest, which occupied 22.17% in RCT1 and 63.16% in RCT2. The corresponding ADR rates were 34.18 and 27.26. The standard deviation (SD) of 7 disease types, which were RA, IgA nephropathy, nephritis, nephrotic syndrome (NS), diabetic nephropathy, psoriasis, lichen and rashes, as well as uterine fibroids, was 8.69 in RCT1. The SD of RA, IgA nephropathy, psoriasis, lichen and rashes was 7.11 in RCT2. It was concluded that the possible ADR distribution ofTripterygium Wilfordiiand its preparations were the highest in the digestive system, reproductive system and liver. Besides, different diseases (i.e., RA, nephritis, NS, and etc.) had huge differences with their correspondent ADR rates. Therefore, it was suggested that specific measures should be taken to select the appropriateTripterygium Wilfordiipreparation, protect the stomach and liver during the application ofTripterygium Wilfordiiand its preparations. During medication, attentions should be paid to the reaction of patients. Stop the medication when necessary to minimize ADR rates to the lowest.
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The study aims to investigate the associations of SUMO4 polymorphisms with tacrolimus concentrations in Chinese renal transplant recipients. Blood samples and clinical data were collected from 132 renal transplant recipients with tacrolimus treatment. CYP3A5*3 genotypes were detected by PCR-RFLP, and SUMO4 (rs237024, rs237025) genotypes were detected by Sequenom® MassARRAY system. SUMO4 rs237024 and rs237025 genotypes were in complete linkage disequilibrium (D' = 1). The dose-adjusted concentration of tacrolimus in SUMO4 rs237024A-rs237025A (GA-GA +AA-AA) carriers was considerably higher than that in GG-GG carriers (P < 0.05). After stratification by CYP3A5*3 genotypes, SUMO4 rs237024A-rs237025A carriers (GA-GA+AA-AA) had a higher dose-adjusted tacrolimus concentration than that in GG carriers in CYP3A5 expresser (P < 0.05). The results illustrated that SUMO4 rs237024 and rs237025 polymorphisms were associated with tacrolimus concentrations, and the test of these genotypes may be useful for individualized medicine of tacrolimus.
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Objective To investigate the effect of clotrimazole on apoptosis of hepatic cells after ischemia-reperfusion injury and its mechanism. Methods Hepatic ischemia-reperfusion rat model was established. Thirty-two male Sprague-Dawley rats were randomly allocated into sham-operated group, model control group, low dose clotrimazole group and high dose clotrimazole group. Apoptosis in hepatic tissue was assessed by TUNEL method. Protein expression levels of CYP3A1,Bcl-2,Bax and PARP were measured by Western blotting. Results As compared with model control group, the apoptosis rate, tissue injury,activity of plasma enzymes and the Bax/Bcl-2 expression ratio were reduced in low and high dose clotrimazole groups. The apoptotic index in both clotrimazole-treated groups was lower than that of model control group with statistically significant difference. CYP3A1 expression was significantly induced by clotrimazole compared to the sham-operated group. Conclusion Clotrimazole may inhibit apoptosis of hepatic cells by up-regulating Bcl-2 and down-regulating Bax, thus produce a protective effect on hepatic ischemia-reperfusion injury and it is also related to the inhibition of PARP shear.
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Objective To discuss the characteristics of pulmonary function in children patients with positive bronchodilating response using impulse oscillation system(IOS) and provide the evidence for clinical diagnosis and performing IOS.Methods Seventy children patients with bronchodilating response using IOS were recruited randomly for this study.Forty children patients with positive bronchodilating response were regarded as positive group and the other thirty children patients with negative bronchodilating response were regarded as negative group.The IOS parameters,for example,total respiratory impedance (Z5),respiratory resistance at 5 Hz(R5),reactance at 5 Hz(X5),peripheral respiratory resistance(Rp) in the two groups were analyzed.Results Z5,R5,X5 of the positive group [(123.45 ± 24.75)％,(120.09 ± 25.40)％,(138.62 ± 33.48)％] were higher than those of the negative group [(103.89 ± 16.30)％,(102.67 ± 18.16)％,(111.85 ± 14.93)％].There were significantly different between the two groups (P ＜ 0.05).Moreover,Rp ≥ 19.89 cmH2O/ (L·s)(1 cmH2O =0.098 kPa) in the positive group was 50.00％(20/40),which was higher than that in the negative group[23.33％(7/30)].There was significant difference between the two groups(P＜ 0.05).Conclusions The impedance of children patients with positive bronchodilating response using IOS is significantly increased,especially X5.When IOS parameters ≥ 100％,especially X5 ≥ 100％,which suggest that perform the bronchodilation test in order to decrease the rate of missed diagnosis of asthma.
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This study was aimed to observe the pharmacodynamics of genistein combined with tripterygium gly-cosides ( GTW ) in the rheumatoid arthritis ( RA ) treatment of ovariectomized rats . SD rats were randomly divid-ed into four groups , which were the sham-operation group , model group , Methotrexate ( MTX ) treatment group ( 2.7 mg/kg ) , and genistein ( 30 mg/kg ) combined with GTW ( 70 mg/kg ) treatment group . There were ten rats in each group . Except the sham-operation group , rats in other groups were ovariectomized and immunized by collagen type II to prepare collagen induced arthritis ( CIA ) model . The intragastric administration of drugs was given once a day for two weeks . The arthritis index , joints swelling degree , and levels of anti-Col II , IL-6 and IL-10 in serum were tested . The results showed that the arthritis index , joints swelling degree , and levels of anti-Col II , IL-6 and IL-10 in serum were all significantly increased in the model group ( P < 0 . 05 or P< 0 . 01 ) . Compared with the model group , the arthritis index and joints swelling degree of both MTX and genistein combined with GTW were significantly decreased . The level of anti-Col II in serum of the MTX treatment group was significantly decreased ( P < 0 . 05 ) . However , there were no significant changes on levels of IL-6 and IL-10 . The levels of anti-Col II and IL-6 in serum of the genistein combined with GTW were significantly decreased ( P < 0 . 05 ) . The level of IL-10 was significantly increased ( P < 0 . 01 ) . It was conclud-ed that genistein combined with GTW can significantly inhibit the pathological progress of CIA on ovariec-tomized rats . Compared with MTX , the genistein combined with GTW received a better regulating effect on the expression of IL-6 and IL-10 . The results suggested that phytoestrogen combined with GTW played a good therapeutic effect on gonadal dysfunction type of RA . It provided novel strategies and experimental evidences on postmenopausal RA treatment in the clinical practice .
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Objective To determine the sequence diversity in groEL gene of Helicobacter pylori isolates,and the antigenicity,immunoreactivity and immunoprotection of recombinant expressed GroEL (rGroEL) against H.pylori-infection in mice.Methods The sequences of groEL genes from H.pylori isolates were detected by PCR and sequencing.A prokaryotic expression system of H.pylori groEL gene using pET42a and E.coli BL21DE3 was generated.SDS-PAGE and Gel Image Analyzer were used to measure the yield of rGroEL and Ni-NTA affinity chromatography was applied to extract the expressed rGroEL.The antigenicity and immunoreactivity of rGroEL were examined using ELISA and Western blot assay.The membrane location of GroEL was determined by ELISA using whole cells of H.pylorias the coated antigen.The immunoprotection of rGroEL was detected in H.pylori-infected mouse model.Results All the 35 tested H.pylori isolates possess the groEL gene with 99.4％ to 100％ sequence identities.The yield of rGroEL expression occupied 56％ of the total bacterial proteins and the extracted rGroEL showed a single band in gel after SDSPAGE.rGroEL could induce the production of high-titer IgG in rabbits or mice.The lgG against whole cell of H.pylori(Hp-IgG) and rGroEL-IgG could recognize as well as combine with rGroEL protein.GroEL is a superficial protein antigen of H.pylori.The immunization with 50,100 or 200 μg rGroEL prevented 50.0％ (6/12),75.0％ (9/12) or 91.7％ (11/12) mice from infection of H.pylori strain SS1.The immunoprotective rate ( 91.7％ ) due to the immunization of 200 μg rG roEL was significantly higher than that ( 50.0％ ) of 50 μg rGroEL ( P＜0.05 ).Conclusion The GroEL protein is a sequence-conserved,immunogenic and immunoprotective superficial antigen of H.pylori,which can be used as the immunogen candidate for developing genetically engineering vaccines.
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ObjectiveTo determine the diversity of outer membrane protein expression of Leptospira interrogans during infection of human THP-1 monocytes,and protein antigen candidates for developing leptospiral genetically engineering vaccines.MethodsThe outer membrane proteins(OMPs) of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai before or after infection of human THP-1 monocyte line were extracted using Triton X-114 method.The OMPs extracts were separated by two-dimensional gel electrophoresis(2-DGE) and then detected using silver staining method.Four up-regulated and four down-regulated OMPs were selected for hydrolysis by trypsin and then identified by using liquid chromatography-tandem mass(LC-MS/MS) method.The transmembrane regions and signal peptides in the eight target OMPs were analyzed using bioinformatic software.Several real-time fluorescent quantitative RT-PCRs were performed to determine the mRNA level changes of the eight target genes of L.interrogans strain Lai before or after infection of THP-1 cells.The prokaryotic expression systems of target genes were generated and the immunoprutection of target recombinant proteins was determined using leptospire-infected guinea pigs model.ResultsAfter a 60 min infection of THP-1 cells,four OMPs( Loa22,GroEL,F0F1 ATP synthase alpha- and beta-subunits) of L interrogans strain Lai were noticeably up-regulated ( P＜0.05 ),while the other four OMPs( FlaB2,LigB,OmpA family protein and OmpA) were significantly down-regulated (P＜0.05).The results of real-time fluorescent quantitative RT-PCRs were coincident with those confirmed by the 2-DGE phus silver staining.The bioinformatic analysis indicated that among the eight OMPs,OmpA and OmpA family protein belonged transmembrane proteins,while the rest had no membrane structure.Loa22,LigB and OmpA family protein contained a signal peptide in their sequences.200 μg of the recombinant proteins rLoa22 and rGroEL presented 75.0％ immunoprotective rates in the infected guinea pigs.Conclusion An obvious change of the OMP expression map of L.interrogans strain Lai occurs during infection of host cells.The up-regulated leptospiral OM Ps during infection,especially for the GroEL and Loa22 proteins,can be used as the immunogen candidates for developing leptospiral genetically engineering vaccines.
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ObjectiveTo evaluate the feasibility of constructing tissue engineering cardiac patch with photooxidationfixed acellular bovine pericardium.MethodsFresh bovine pericardia were treated by dye-mediated photooxidation after decellularization.Some of them were seeded with bone marrow stromal cells(MSCs) isolated from male SD rats to construct cardiac patches.Myocardial infarction(MI) model was made in female SD rats by left anterior descending coronary ligation(LAD).One week later, the confirmed MI rats were divided into three groups randomly, group MI (n = 15)without any treatment; group P (n = 18) with photooxidated pericardia implantation ; group P + C (n = 18) with seeded pericardia implantation.A sham group (n = 10) was also performed with opening and closing chest twice only.The heart were explanted at 2 or 4 weeks after implantation, and examined histologically and immunohistochemically.The heart function was evaluated by echocardiography at 4 weeks before excising the rats.ResultsThere were no cells or cell debris remained in bovine pericardium tissue.The fiber structure became condensed after photooxidation.The seeded cells formed a continuous layer on the surface of the tissue.The pericardial degradation level and newly formed microvessel density were larger in group P + C than in group P after 2 [ (13.7 ±5.2)个/mm2 vs (7.1 ±3.1)个/mm2, P＜0.05]and4 [(22.6 ±4.9)个/mrn2 vs (14.1 ±5.3)个/mm2, P＜0.05]weeks.Four weeks after transplantation, cardiac echocardiography showed left ventricular ejection fraction(LVEF) was lower in group MI (44.8 ± 4.4) ％ and group P (48.4 ± 5.0) ％ compared with group P + C (49.3 ± 4.8) ％, left ventricular fractional shorterning(LVFS) was lower in group MI (18.0 ± 2.2) ％ and group P (19.8 ± 2.5) ％ compared with group P + C (20.4 ±2.5) ％, the difference between P + C and MI was significant.ConclusionTransplantation of the tissue engineered bovine pericardial patches with dye-mediated photooxidation can improve heart function in MI rats.This kind of patches demonstrates a promising prospect in the future.
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Objective To investigate the expression of 5-lipoxygenase (5-LOX) in endometriosis and analyze the relationship of 5-LOX and angionesis in endometriosis. Methods Immunochemical method was used to determine the expression of 5-LOX, VEGF, and the value of MVD in ectopic endometrium (ec-topic group) and eutopic endometrium (eutopic group) of 32 patients with EMs and in eutopic endometrium ( control group) of 25 patients without EMs. Result The expressions of 5-LOX and VECF and the value of MVD in ectopic groups were 0. 43 ± 0.09,0. 39 ±0.07 and 0.40 ± 0.09, respectively, which were higher than that of eutopic group and control group. The expressions of 5-LOX and VEGF and the value of MVD in eutopic group were 0. 35 ± 0. 08, 0.30 ± 0. 06 and 0. 33 ± 0. 08, respectively, which were higher than that of control group ( P <0.05). In ectopic group, the expressions of 5-LOX and VEGF, and the value of MVD in stage III/IV, were much higher than that of stage I/II ( P <0. 05); the expressions of 5-LOX and VEGF and the value of MVD were related to the severe of endometriosis through Spearman correlation analysis. The expressions of 5-LOX were shown to be significantly related to the expressions of VEGF and the value of MVD in ectopic and eutopic groups ( P <0. 05). Conclusion There is a high expression of 5-LOX in endometriosis, suggesting that 5-LOX may be participate in the pathogenesis of endometriosis. 5-LOX may promote angiogenesis of EMs through up - regulating the expression of VEGF.
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OBJECTIVE To investigate the causes of fever and risk factors in portal hypertensive patients after combined operation(devascularization+shunt).METHODS Forty five cases of portal hypertension(PHT) after combined operation were retrospectively and prospectively analyzed.RESULTS Complications caused 88% post operational fever.The most common cause was hydrothorax,hematocele or hydrops and infection in splenic recess.Long-term fever was related to liver function(P