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Objective:To obtain hepatitis B virus capsid-like particles (CLPs) displaying B-cell linear epitopes of SARS-CoV-2 spike protein and evaluate their immunogenicity.Methods:Four recombinant plasmids expressing fusion proteins (M1-HBc, S1-57-HBc, S14P5-HBc and S21P2-HBc) were constructed by separately replacing codon of alanine at position 80 of hepatitis B virus core protein (HBc) with four genes coding for four B-cell linear epitopes (M1, S1-57, S14P5 and S21P2). These four recombinant proteins were expressed in E. coli BL21 Star (DE3) strains. The expression products were identified using SDS-PAGE, Western blot and native agarose gel electrophoresis (NAGE). CLPs were purified by sucrose density gradient ultracentrifugation, verified for antigenicity by Western blot and used to immunize BALB/c mice. Serum antibody titers were detected by ELISA. Results:The recombinant fusion proteins M1-HBc and S1-57-HBc self-assembled into M1-CLP and S1-57-CLP. The titer of antibody against S1-57 polypeptide in S1-57-CLP-immunized mouse serum approached 1∶1 000 000.Conclusions:Hepatitis B virus CLPs displaying SARS-CoV-2 M1 or S1-57 linear epitopes are successfully expressed in a prokaryotic system and purified. S1-57-CLP has good immunogenicity. This study provides a new idea for the development of novel diagnostic reagents and vaccines for SARS-CoV-2.
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Smart manufacturing still remains critical challenges for pharmaceutical manufacturing. Here, an original data-driven engineering framework was proposed to tackle the challenges. Firstly, from sporadic indicators to five kinds of systematic quality characteristics, nearly 2,000,000 real-world data points were successively characterized from Ginkgo Folium tablet manufacturing. Then, from simplex to the multivariate system, the digital process capability diagnosis strategy was proposed by multivariate Cpk integrated Bootstrap-t. The Cpk of Ginkgo Folium extracts, granules, and tablets were discovered, which was 0.59, 0.42, and 0.78, respectively, indicating a relatively weak process capability, especially in granulating. Furthermore, the quality traceability was discovered from unit to end-to-end analysis, which decreased from 2.17 to 1.73. This further proved that attention should be paid to granulating to improve the quality characteristic. In conclusion, this paper provided a data-driven engineering strategy empowering industrial innovation to face the challenge of smart pharmaceutical manufacturing.
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Objective:To explore the relationship between constitutional types of Chinese medicine and Alzheimer's disease (AD) and to construct an early warning model for AD risk.Methods:In the established multimodal interventions to delay dementia and disability in rural China (MIND-China) study, 4 033 elderly subjects aged ≥60 years old were included. The data including demographic, underlying disease and neuropsychological data were collected.The Chinese medicine service record form for the elderly was used to assess constitutional types of Chinese medicine and to apply the NIA-AA diagnostic criteria published by the National Institute on Aging and the Alzheimer's Association in 2011 for the diagnosis of clinically likely AD. Logistic regression analysis and AD risk prediction models were constructed using R statistical software, and the final prediction results were presented using columnar plots.Results:The MIND-China cohort was dominated by the abnormal constitution (69.28%), of which Phlegm-wetness type was the most common (58.05%), followed by Yang-deficiency type (23.85%). The most constitutional type of Chinese medicine among AD patients was Phlegm-wetness type (54.35%), followed by Qi-depression type (38.04%). Multi-factorial logistic regression analysis suggested that increasing age ( β=0.101, P<0.001, OR=1.107, 95% CI=1.069-1.146) and Qi-depression type ( β=0.622, P=0.016, OR=1.862, 95% CI=1.116-3.076) were able to increase the risk of developing AD, while education ( β=-1.047, P<0.001, OR=0.351, 95% CI=0.205-0.584) was able to reduce the risk of developing AD. By using the risk score model to calculate the total risk score for each subject and plotting the receiver operating characteristic curve (ROC), the area under the ROC was 0.769 and the calibration curve showed excellent consistency between prediction and reality. Conclusion:Older adults with Qi-depression type are significantly associated with an increased likelihood of AD.
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@#Objective To investigate the effect of enteral nutritional support beginning at different time on aspiration events and immune function in the early post-traumatic (within 14 days) period in patients with severe traumatic brain injury.Methods From June, 2018, to February, 2021, 75 patients with acute traumatic brain injury admitted in the Department of Neurosurgery of the Second People's Hospital of Lianyungang were randomly divided into early feeding group (24 to 48 hours, n=35) and delayed feeding group (48 to 120 hours, n=40). The 14-day reflux rate, aspiration rate, incidence of aspiration pneumonia, immune indexes and complications were compared between two groups.Results There was no significant difference in the reflux rate, aspiration rate and the incidence of aspiration pneumonia between the two groups (P > 0.05). The levels of IgG, IgA, and complements C3 and C4 were significantly higher in the early feeding group than in the delayed feeding group (|t| > 1.720, P<0.001). The incidence of non-aspiratory lung infections was significantly lower in the early feeding group than in the delayed feeding group (χ2=4.728, P<0.05).Conclusion Initiating enteral nutrition within 24 to 48 hours after injury may preserve immune function and reduce the incidence of non-aspiratory lung infections in patients with severe traumatic brain injury.
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Objective:Through TTC staining, immunohistochemical analysis, RT-PCR and hind limb motor function evaluation and other experimental methods, to explore the regulatory mechanism of metformin on anti-apoptosis in rats with spinal cord injury (SCI).Methods:Establish a rat spinal cord injury model. Through Basso-Beattie -Bresnahan locomotor rating scale (BBB) and cant test to evaluate the recovery of hindlimb motor function in rats. The changes of necrotic area of spinal cord tissue were compared by TTC staining. Extraction of rat spinal cord tissue, by Dot blot analysis and immunohistochemical detection of the hydroxyl of DNA methylation level. By qPCR, Western Blot detection TET2mRNA and protein expression level, and the changes in the scope of spinal cord injury were detected by inhibiting the expression of TET2. The interaction between TET2 and Foxo3a was detected by immunoblotting and immunoprecipitation. Through RT-PCR assay Foxo3a downstream related changes in the level of gene expression.Results:Compared with the SCI+NS group, the necrotic area of the spinal cord tissue was reduced after metformin treatment, and the BBB score and the incline test score were higher ( P<0.05). At the same time, we found that the levels of TET2mRNA and protein increased significantly after SCI at 24 h, and the 5-hmC level of DNA increased. The levels of TET2mRNA and protein and 5-hmC increased further after the use of metformin. After using SC-1, compared with the SCI+MET group, the level of 5-hmC decreased and the area of infarction increased. After SCI, the mRNA levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the mRNA levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). After SCI, the 5-hmC levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the 5-hmC levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). Conclusion:Metformin can promote the interaction between TET2 and Foxo3a, increase the 5-hmC level of the overall DNA, and inhibit the activation of related apoptosis genes, thereby improving tissue damage and nerve function recovery after spinal cord injury.
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Objective To evaluate the biomechanical effects of lateral meniscus posterior root (LMPR) tears fixed at different suture positions, so as to investigate the optimal suture method for repairing LMPR tears. Methods Eight fresh cadaveric knees were used. Each knee was tested under 6 conditions: intact knee, ruptur of LMPR, suture of LMPR to the center point of root insertion, suture of LMPR posterior, interior and later 5 mm to the center point of root insertion, respectively. The peak contact pressure, the average contact pressure and contact area were evaluated using a Tek-scan sensor positioned between the meniscus and tibial plateau, under 1 kN compressive loading, at 0 degree knee extension. Results In the lateral compartment, the average contact pressure and peak pressure significantly increased under rupture of LMPR compared with the intact state (P0.05). Conclusions The average contact pressure, the peak contact pressure and the contact area between the lateral meniscus and the tibial plateau changed obviously due to the LMPR tears. When the suture position was 5 mm lateral to the center point of root insertion, similar biomechanical function with the intact knee could be obtained.
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Objective@#To investigate the effect of an ubiquitin-like modification NEDDylation on HBV life cycle.@*Methods@#Cells were incubated with various concentrations of MLN4924, which is a selective inhibitor of neural precursor cell-expressed developmentally down regulated 8 (NEDD8) activating enzyme (NAE), to block cellular protein NEDDylation. And a series concentration of lamivudine (LAM), an inhibitor of HBV polymerase was set as control. Then we tested HBV DNA replication, protein expression and nucleocapsids formation. Huh-7 cells were transfected with polymerase-inactivated HBV expression plasmid, then incubated with 0.1 μmol/L MLN4924 or çvehicle-DMSO. Then we tested capsids and capsid-associated RNA.@*Results@#MLN4924 inhibited HBV replication in a dose-dependent manner (0~2 μmol/L). When the concentration of MLN4924 was 0.1 μmol/L, the amount of nucleocapsids in the cells did not change significantly, however the encapsidation of HBV pregenomic RNA (pgRNA) decreased by about 47%.@*Conclusions@#NEDDylation is involved in hepatitis B virus pregenomic RNA encapsidation.
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Cerebral small vessel disease is the common clinical,imaging and pathological syndrome that contribute to the onset and progression of cognitive impairment,and it also plays a crucial role on stroke,dementia,brain aging,and neurodegenerative disease.Cerebral small vessel disease can be detected and diagnosed using structural brain magnetic resonance imaging (MRI).The main imaging features of various cerebral small vessel disease include small subcortical infarcts,lacunes of presumed vascular origin,white matter hyperintensities,enlarged perivascular spaces,cerebral microbleeds,and global and regional brain atrophy.Other emerging imaging features are detectable at very high field strength MRI include microinfarcts.Cerebral small vessel disease used to be considered clinically silent.However,in the past decades or so,numerous studies have investigated the association between various imaging markers of cerebral small vessel disease and cognitive dysfunction (e.g.vascular cognitive impairment and Alzheimer's disease).The results have shown that these neuroimaging markers are associated with cognitive dysfunction,and can be used to predict the risk of dementia and the progression of cognitive impairment.This review article seeks to summarize neuroimaging studies that examine the associations between various markers of cerebral small vessel disease and cognitive impairment.
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Objective To explore the early clinical effects of lumbar discectomy associated with annulus repair in the treatment of lumbar disc herniation.Methods A prospective study was conducted to review 56 patients with lumbar disc herniation who accepted discectomy surgery in Subei People's Hospital of Jiangsu Province from January 2014 to September 2015,including 28 cases of discectomy associated with annulus repair (repair group) and 28 cases of discectomy (control group).Oswestry disability index and visual analog scale scores were recorded.Simultaneously,incision length,operative time,blood loss,hospitalization time,surgical complications,and postoperative recurrence of lumbar disc herniation were recorded.Results All patients completed the follow-up for 12 to 18 months (14.5 ± 1.3).There was no difference between the repair and control groups in the incision length,blood loss and hospitalization time (P ＞ 0.05).The operative time of the repair group was longer than that of the control group,but the difference was not statistically significant (P ＞ 0.05).The Oswestry disability index and visual analog scale scores for lumbar and lower limb pain significantly decreased in both groups after surgery (P ＜ 0.05).The visual analog scale scores at 24 hours and 3 days after surgery in the repair group were less than that in the control group (P ＜ 0.05).The satisfactory rate of treatment in the repair group was slightly higher than that in the control group,but the difference was not statistically significant (P ＞ 0.05).There was no recurrence in the repair group,but 2 recurrence cases in the control group (P ＞ 0.05).Conclusions These findings indicate that discectomy associated with annulus repair is a safe and reliable method to obtain remarkable early clinical results and can reduce the recurrent rate in the treatment of lumbar disc herniation.
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OBJECTIVE:To optimize the dry granulation technology conditions for Yinqiao baidu tablet. METHODS:Using granulating difficulty degree and disintegration time as investigation indexes,ratio and amount of accessories microcrystalline cellu-lose and compressible starch in Yinqiao baidu tablet,moisture content of the sprayed powder were screened. Using yield of particle and angle of repose as indexes,L9(34)orthogonal test was used to optimize the wheel pressure,rotating speed and feeding speed in dry granulation technology,and verification test was conducted. RESULTS:The ratio of microcrystalline cellulose and compress-ible starch was 7:3,and mixing ratio of the two with spray powder+inclusion compound was 1:5. The moisture content of spray powder was controlled in 1％-2％. The optimal technology was as follow as wheel pressure of 3.5 MPa,roller speed of 4 r/min and feeding speed of 10 r/min. In verification test,average yield of particle was 69.2％ and angle of repose was 31.5 °. Transfer rate of chlorogenic acid had reached over 92％,and RSD of each index was below 2.53％(n=3). CONCLUSIONS:Each index of parti-cle prepared by optimized accessories formulation and technology shows good reproducibility and feasibility,and the technology is stable and suitable for production.
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Objective To explore the expression of CD44v8 protein in human bladder and urothelial carcinoma ,as well as the value in diagnosis of human bladder and urothelial carcinoma .Methods RT‐PCR was used to analysis the expression of CD44v8 protein in 75 patients with bladder and urothelial carcinoma in the pathological stage and clinical stage and 20 subjects of normal bladder mucosa were collected as control .Results CD44v8 protein was negative in all normal bladder and urothelial mucosa ,the copy number was less than 1 × 102 copy/mL ;26 cases of bladder and urothelial carcinoma was positive ,and the positive rate of CD44v8 was 34 .7% ,and Ct values were less than 35 and copy number was greater than 1 × 104 copy/mL .Positive rate was correla‐ted with high pathological grades and TNM stages ,but no significant difference was observed in recurrence of tumor .Conclusion CD44v8 could be useful indicator for the assessment of pathological grades and TNM stages of bladder and urothelial carcinoma .
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Objective To investigate the expression of CD44 varant 2(CD44v2) in bladder and urothelial carcinoma ,and to study its significance in the diagnosis of human bladder and urothelial carcinoma .Methods Real‐time fluorescent quantitative PCR was used to analyze the expression of CD44v2 protein in 70 bladder and urothelial carcinoma tissue samples from patients in different pathological and clinical stages .Meanwhile ,20 tissue samples of normal bladder mucosa were collected as controls .Results CD44v2 expression was negative in normal bladder and urothelial mucosa ,the gene copies were less than 1 × 102 copies/mL ,while the posi‐tive expression rate of CD44v2 was 42 .9% (30/70) ,and Ct values were less than 35 and copy number was greater than 1 × 104 cop‐ies/mL .Positive expression rate was correlated with high pathological grades and TNM stages .Conclusion CD44v2 could be an useful indicator for the early assessment of bladder and urothelial carcinoma .
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This study was aimed to optimize the extraction technology of Shu-Feng Ding-Chuan(SFDC) granules in the Huang-Qin(HQ)group by Box-Behnken response surface methodology(RSM).With three major characteristic components (baicalin,praeruptorin A and solid content in extraction liquid),the effects of four factors,such as the concentration of ethanol,the dosage of ethanol,the duration of extraction and the extraction times,were investigated by the single factor experiment.Then,the range of parameters of key factors was further studied and explored by Box-Behnken RSM.The results of single factor experiment and Box-Behnken design showed that the optimum preparation conditionswere 8-fold 70% ethanol,extracted for 2 times,with 1.0 h per each time.Under these conditions,the transfer rates of baicalin and praeruptor in A were 89.57% and 87.90%,respectively.And the transfer rateof solid content was 32.35%.It was concluded that the single factor experiment combined with RSM can be used in theoptimization of extraction technology for SFDC granules in the HQ group.This technique wasstable and feasible,which provided scientific evidences for the industrial production.
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Objective:To investigate the relationship between cancer-related fatigue and cortisol in cancer patients and elucidate the underlying mechanism. Methods:A total of 80 cancer cases were divided into two groups:fatigue group (50 cases with cancer-related fatigue) and non-fatigue group (30 cases without fatigue). The scores were evaluated through the Multidimensional Fatigue Symptom Inventory-Short Form (MFSI-SF) and the Fatigue Symptom Inventory (FSI) report. Serum specimens were examined through electrochemiluminesence immunoassay and enzyme-linked immunosorbent assay. Serum cholesterol was examined through the CHOD-PAP method, and serum total protein and albumin were determined via the Biuret method. Agarose gel electrophoresis was conducted to determine alpha 2 globulin ratio and to calculate serum alpha 2 globulin concentration. Results: The cortisol level in the fatigue group was significantly lower than that in the non-fatigue group[(119.68±5.34) nmol/L vs. (163.45± 31.49) nmol/L, P<0.05], and the adrenocorticotropic hormone (ACTH) level in the fatigue group was significantly higher than that in the non-fatigue group [(104.50 ± 17.15) ng/L vs. (51.43±13.24) ng/L, P<0.05]. Cortisol negatively correlated with MFSI-SF (r=-0.867, P<0.001) but positively correlated with ACTH (r=0.809, P<0.001). Furthermore, cortisol negatively correlated with FSI (r=-0.747, P<0.001) but positively correlated with ACTH (r=0.70, P<0.001). The levels of serum cholesterol [(1.25±0.70) mmol/L vs. (3.28±0.73) mmol/L, P<0.05], albumin[(18.24 ± 7.03) g/L vs. (37.40 ± 8.05) g/L, P<0.05], and alpha-2 globulin [(2.25±1.07) g/L vs. (5.36±1.09) g/L, P<0.05]were significantly lower in the fatigue group than in the non-fatigue group. Conclusion:The patients with cancer-related fatigue exhibited increased MFSI-SF score, decreased serum cortisol level, and enhanced ACTH level. The low serum cortisol levels caused a disorder in the serum ACTH and cancer-related fatigue of malignant tumor patients. The mechanism underlying the reduction in serum cortisol level correlated with the insufficient amounts of serum cholesterol, the composite material of cortisols, and of serum albumin, particularly alpha-2 globulin, the carrier protein of serum cholesterol.
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This study was aimed to acquire the dynamic changes of chlorogenic acid and luteoloside of honeysuckle at different collecting periods in order to decide the best harvesting time of honeysuckle in Donghai County, Jiangsu Province. The content determination method used in the detection of chlorogenic acid and luteoloside of honeysuckle was from the 2010 edition of the Chinese Pharmacopoeia. The skills of HPLC fingerprint characteristic features, and the yield of pressed flowers were combined in the evaluation of honeysuckle at the three-green period, two-white pe-riod, great-white period, silver-flower period and the golden-flower period. The results showed that the content of honeysuckle at different blossoming stages had obvious changes in content of chlorogenic acid and luteoloside, as well as the pressed flower quality and yield of the flower. It was concluded that the optimal harvest time of honey-suckle was for the two-white period and the great-white period, which was consistent with the real origin.
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Objective To establish a simple and stable orthotopic segmental small bowel transplantation model in rats.Methods Eighty male SD rats were divided into donors and recipients.Orthotopic segmental small bowel transplantation was performed by end-to-side anastomosis between donor abdominal aorta tundish-shape patch with arteria mesenterica superior pedicle and abdominal aorta of the recipients; end-to-end anastomosis between the portal vein of the donors and the left renal vein of the recipients was done using the cuff technique ; the large part of the small bowel of the recipients was excised,and it was replaced by the segmental intestine of the donors.Results The operation time of the donors and recipients were (40 ± 5) minutes and (50 ±8)minutes,respectively.The warm ischemia time and cold ischemia time were (5 ± 2) minutes and (15 ± 5) minutes,respectively.The anastomosis time of arteries and veins were (5 ± 2) minutes and (4 ± 2) minutes,respectively.The survival time of 90.0％ (36/40) of rats was more than 10 days.Conclusion The modified rat model of orthotopic segmental small bowel transplantation is easy to manipulate,and has the advantages of short operation time,high survival rate and stability.
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ObjectiveTo Summarize the experience of totally thoracoscopic operation for atrial myxoma.Methods From December 2007 to November 2011,44 patients with atrial myxoma,including 37 cases of Left atrial myxoma,6 cases of Right atrial myxoma,1 case of double atrial myxoma were treated totally under thoracoscope.Surgical procedures were performed through 3 troears inserted at the right chest wall,and catheters were placed in the right femoral artery and vein (or in the right atrium,femoral vein)to set up extracorporeal circulation.The ascending aorta was cross-clamped with long tailor-made forceps and the myocardium was protected by coronary perfusion with cold crystalloid (blood) cardioplegia.ResultsAll the operations were completed successfully.The mean extracorporeal circulation and cross-clamping time were (49 ± 18 )min and (28 ± 10) min respectively.Postoperative ventilation was withdrawn in(3.8 ± 1.4) h,and the patients were discharged from the hospital in (6.8 ± 1.3 ) d.3 of the patients had postoperative complications,including 1 case of fat liquefaction of the incision at the right groin ( delayed healing),2 cases of subcutaneous emphysema ( healed by bandaged chest).No severe complications occurred in this series.UCG performed 3 - 5 days after the operation revealed surgical results were satisfactory.Followup up to 2 months to 4 years were available in all cases.During the period,the heart function was confirmed as level Ⅰ.ConclusionVideo-assisted thoraeoseopie cardiac surgery is feasible,safe,and minimal invasive for patients,resulting in quick recovery and good cosmetic outcomes.
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ObjectiveTo summarize the experience of totally thoracoscopic operation for congenital heart diseases.MethodsFrom September 2000 to November 2011,1281 patients with congenital heart disease,including 1016 cases of atrial septal defect,110 cases of ventricular septal defect,61 cases of atrioventricular tube defects,33 cases of tetralogy of Fallot,24 cases of part anomalous pulmonary venous connection,12 cases of pulmonary valve stenosis,9 cases of patent ductus arteriosus,8 case of triatriatum,7 cases of unroofed coronary sinus syndrome,and 1 case of total anomalous pulmonary venous connection were treated totally under thoracoscope.Surgical procedures were performed through 3 troears inserted at the right chest wall,and catheters were placed in the right femoral artery and vein ( or in the right atrium,femoral vein) to set up extracorporeal circulation.The ascending aorta was cross-clamped with long tailor-made forceps and the myocardium was protected by coronary perfusion with cold crystalloid (blood) cardioplegia.ResultsAll the operations were completed successfully.The mean extracorporeal circulation and cross-clamping time were ( 42 ± 16 ) min and ( 21 ± 9 ) min respectively.Postoperative ventilation was withdrawn in(4.1 ± 1.5 ) h,and the patients were discharged from the hospital in(7.1 ± 1.4) d.35 of the patients had postoperative complications,including 16 cases of right pneumothorax (healed by thoracentesis),12 cases of subcutaneous emphysema ( healed by bandaged chest) and 7 cases of fat liquefaction of the incision at the right axillary( 3 cases) and groin(4 cases) ( delayed healing).No severe complications occurred in this series.UCG performed 4 - 8 days after the operation revealed no residual shunt.Follow-up up to 3 months to 9 years were available in 914 cases.During the period,the heart function was confirmed as level Ⅰ - Ⅱ.ConclusionTotally thoracoscopic cardiac surgery is feasible,safe,and minimal invasive for patients,resulting in quick recovery and good cosmetic outcomes.
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Objective To observe the modulatory effects of NF-κB on dynamic changes of TNF-α、IL-1β and ICAM-1 in rats with severe acute pancreatitis (SAP),and to assess the effects of pretreatment with mycophenolate mofetil (MMF) on them.Methods A total of 104 male Wistar rats were randomly divided into normal group,sham operation group,SAP group and MMF group.(1) In normal group,rats had just an incision on abdomen wall and sutured (n =8) ; (2) In sham operation group,rats underwent laparotomy with flipping over the pancreas and gendy striking five times by using blunt instrument and abdomen was closed (n =32); (3) In SAP group,SAP in rats was induced by the retrograde injection of sodium taurocholate (STC) into the pancreatic duct (n =32) ; (4) In MMF group,MMF (250mg/kg)was injected intraperitoneally one hours after injection of STC (n =32).Rats were saerificed by abdominalaorta exsanguination separately at intervals of 3,6,12 and 24 h after modeling of pancreatitis.Histologic score of pancreatic tissue was assessed,and serum amylase (AMS) and serum C-reactive protein (CRP)were measured by automatic biochemical detector.The serum levels of inflammatory cyctokines (TNF-α,1L-1β and 1CAM-1 ) were measured by using ELISA,and expression of NF-κB mRNA in pancreatic tissue was measured by using quantitative PCR.Results Compared with sham operation group,histologic score of pancreatic tissue,serum amylase,TNF-α、IL-1β and ICAM-1 in SAP group were increased significantly (P ＜ 0.05 ).Pretreatment with MMF obviously deceased histologic score,and the serum levels of amylase,TNF-α,IL-1β and ICAM-1 and the expression of NF-κB mRNA were higher than those in SAP group in early stage (P ＜0.05 ).Conclusions Cytokines play an important role in early stage of SAP.MMF could attenuate the severity of inflammatory response through inhibiting cellular immunity and modulating the proinflammatory cytokines produced.
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<p><b>OBJECTIVE</b>Applying for the activity of enzyme in vitro,the research optimized the best preparation procedure for the anticoagulated blood region from Lumbricus.</p><p><b>METHOD</b>All through our experiment, the content of protein and theactivity of enzyme were examined. The extraction process, the refining technology, concentration processes of Lumbricus were optimized with single factor checking and orthogonal design method.</p><p><b>RESULT</b>At 37 degrees C, the coarse powder of Lumbricus soaking with 15 fold of 0.9% sodium chloride and ultrasonic extracting 40 minites for three times was the best ultrasonic extraction. Utrafiltration membrane with molecular weights of 30 x 10(3) for refining and 10 x 10(3) for concentrating were selected.</p><p><b>CONCLUSION</b>Ultrasonic extraction and membrane separation technology, to well improve the effect of purification for the anticoagulant site of Lumbricus, is conducive to further study.</p>