Effects of poly(A) tails with different lengths on mRNA expression in vitro and stability of transcription template in Escherichia coli / 中华微生物学和免疫学杂志

Objective:To investigate the effects of poly(A) tails with different lengths on mRNA expression in vitro and the passage stability of transcription template with poly (A) tail in Escherichia coli ( E. coli). Methods:Plasmids with poly(A) tails of 38, 60, 103, 125 and 126 (60 nt+ 6 nt spacer+ 60 nt) nt were designed and constructed. Then the plasmids were linearized by single enzyme digestion and used as transcription template for preparing enhanced green fluorescent protein (EGFP)-mRNA. EGFP-mRNA containing poly(A) tails of different lengths were transfected into 293T cells and the expression of EGFP was detected by flow cytometry. As to stability test, the template plasmids with poly (A) tail of 125 and 126 nt were transformed into E. coli TransStbl3 and Top10 competent cells. Seven clones were selected for culture and plasmid extraction, and then the plasmids were digested by restriction enzyme and detected by capillary electrophoresis. For passage stability, three correctly sequenced clones of each group were selected for continuous passage at 37℃, and the plasmids were extracted and digested every two generations for capillary electrophoresis. At the same time, the correctly sequenced clones of 125 nt group were also passaged at 30℃, and the plasmids were also extracted and digested every two generations for capillary electrophoresis. Results:The transcription templates with poly(A) tail of different lengths were successfully constructed. Flow cytometry showed that the fluorescence expression of the template plasmids with poly (A) tail of 103 and 125 nt were significantly higher than that of 38 and 60 nt. The fluorescence expression of the plasmid with poly (A) tail of 126 nt was significantly higher than that of all other groups. The percentages of stable sequences of the template plasmid with poly(A) tail of 125 nt in TransStbl3 and Top10 competent cells were 76% and 91%, respectively. The results of continuous passage showed that poly(A) tail of 125 nt could be stable to the 4th generation at 37℃ in both TransStbl3 and Top10 competent cells, and stable to the 16th and 10th generations at 30℃. The percentages of stable sequences of the template plasmid with poly(A) tail of 126 nt in TransStbl3 and Top10 competent cells were 95% and 48%, respectively. The results of continuous passage showed that poly(A) tail of 126 nt could be stable to the 12th generation at 37℃ in both TransStbl3 and Top10 competent cells.Conclusions:The length and composition of poly(A) tail in mRNA affected the expression of target protein. Adding a spacer with a length of 6 nt to poly(A) tail and low temperature culture were both helpful to improve the stability of the template plasmid, which provided a reference for the design and preparation of in vitro transcription template of mRNA vaccine.

Development of auxiliary substation system for endoscopic ultrasound bile duct scanning based on deep learning / 中华消化内镜杂志

Objective:To construct a deep learning-based artificial intelligence endoscopic ultrasound (EUS) bile duct scanning substation system to assist endoscopists in learning multi-station imaging and improve their operation skills.Methods:A total of 522 EUS videos in Renmin Hospital of Wuhan University and Wuhan Union Hospital from May 2016 to October 2020 were collected, and images were captured from these videos, including 3 000 white light images and 31 003 EUS images from Renmin Hospital of Wuhan University, and 799 EUS images from Wuhan Union Hospital. The pictures were divided into training set and test set in the EUS bile duct scanning system. The system included filtering model of white light gastroscopy images (model 1), distinguishing model of standard station images and non-standard station images (model 2) and substation model of EUS bile duct scanning standard images (model 3), which were used to classify the standard images into liver window, stomach window, duodenal bulb window, and duodenal descending window. Then 110 pictures were randomly selected from the test set for a man-machine competition to compare the accuracy of multi-station imaging by experts, advanced endoscopists and the artificial intelligence model.Results:The accuracies of model 1 and model 2 were 100.00% (1 200/1 200) and 93.36% (2 938/3 147) respectively. Those of model 3 on the internal validation dataset in each classification were 97.23% (1 687/1 735) in liver window, 96.89% (1 681/1 735) in stomach window, 98.73% (1 713/1 735) in duodenal bulb window, and 97.18% (1 686/1 735) in duodenal descending window. And those on the external validation dataset were 89.61% (716/799) in liver window, 92.74% (741/799) in stomach window, 90.11% (720/799) in duodenal bulb window, and 92.24% (737/799) in duodenal descending window. In the man-machine competition, the accuracy of the substation model was 89.09% (98/110), which was higher than that of senior endoscopists [85.45% (94/110), 74.55% (82/110), and 85.45% (94/110)] and close to the level of experts [92.73% (102/110) and 90.00% (99/110)].Conclusion:The deep learning-based EUS bile duct scanning system constructed in the current study can assist endoscopists to perform standard multi-station scanning in real time more accurately and improve the completeness and quality of EUS.

Prokaryotic expression and serological analysis of the head domain of influenza A virus hemagglutinin / 中华微生物学和免疫学杂志

Objective:To express the head domain of influenza A virus hemagglutinin (HA) in a prokaryotic expression system and to evaluate its immunogenicity.Methods:The genes encoding the HA head domains of H1N1 and H3N2 influenza viruses were cloned into pET-22b(+ ) prokaryotic expression plasmid. After the induction with IPTG, the fusion proteins rH1N1-HA and rH3N2-HA containing HA head domain and His-tag were expressed and obtained from E. coli BL21. SDS-PAGE and Western blot was used to verify the expression of the recombinant proteins. Rabbits were immunized with multiple doses of the purified recombinant proteins to obtain polyclonal antibodies against the HA head domains of H1N1 and H3N2. The immunogenicity of the recombinant proteins was evaluated in BALB/c mice. Results:rH1N1-HA and rH3N2-HA induced protective antibodies (geometric mean titer ≥40) in mice and could be used as protective antigens. Polyclonal antibodies against rH1N1-HA and rH3N2-HA could be used as important materials for Western blot, ELISA and other immunological assays.Conclusions:The HA head domains prepared in this study could be used as protective antigens to induce protective antibodies in mice. Polyclonal antibodies against the HA head domains could be used for immunological and serological studies of influenza A viruses.

Isolation of Fusarium and identification of its toxins from tuberous root of Pseudostellaria heterophylla / 中国中药杂志

Fusarium is the major pathogen of root rot of Pseudostellaria heterophylla. This study aims to explain the possible distribution of Fusarium species and the contamination of its toxin-chemotypes in tuberous root of P. heterophylla. A total of 89 strains of fungi were isolated from the tuberous root of P. heterophylla. Among them, 29 strains were identified as Fusarium by ITS2 sequence, accounting for 32.5%. They were identified as five species of F. avenaceum, F. tricinctum, F. fujikuroi, F. oxysporum, and F. graminearum based on β-Tubulin and EF-1α genes. LC-MS/MS detected 18, 1, and 5 strains able to produce ZEN, DON, and T2, which accounted for 62.1%, 3.4%, and 17.2%, respectively. Strain JK3-3 can produce ZEN, DON, and T2, while strains BH1-4-1, BH6-5, and BH16-2 can produce ZEN and T2. PCR detected six key synthase genes of Tri1, Tri7, Tri8, Tri13, PKS14, and PKS13 in strain JK3-3, which synthesized three toxins of ZEN, DON, and T2. Four key synthase genes of Tri8, Tri13, PKS14, and PKS13 were detected in strains BH1-4-1, BH6-5, and BH16-2, which were responsible for the synthesis of ZEN and T2. The results showed that the key genes of toxin biosynthesis were highly correlated with the toxins produced by Fusarium, and the biosynthesis of toxin was strictly controlled by the genetic information of the strain. This study provides a data basis for the targeted prevention and control of exo-genous mycotoxins in P. heterophylla and a possibility for the development of PCR for rapid detection of toxin contamination.

Screening of zearalenone-degrading bacteria and analysis of degradation conditions / 中国中药杂志

Zearalenone(ZEN) is a mycotoxin produced by Fusarium, possessing estrogen-like effects, carcinogenicity, and multiple toxicities. To seek more efficient and practical agents for biological detoxification and broaden their application, this study isolated 194 bacterial strains from the moldy tuberous root of Pseudostellaria heterophylla, which were co-cultured with ZEN. An efficient ZEN-degrading strain H4-3-C1 was screened out by HPLC and identified as Acinetobacter calcoaceticus by morphological observation and molecular identification. The effects of culture medium, inoculation dose, culture time, pH, and temperature on the degradation of ZEN by H4-3-C1 strain were investigated. The mechanism of ZEN degradation and the degrading effect in Coicis Semen were discussed. The degradation rate of 5 μg·mL~(-1) ZEN by H4-3-C1 strain was 85.77% in the LB medium(pH 6) at 28 ℃/180 r·min~(-1) for 24 h with the inoculation dose of 1%. The degradation rate of ZEN in the supernatant of strain culture was higher than that in the intracellular fluid and thalli. The strain was inferred to secret extracellular enzymes to degrade ZEN. In addition, the H4-3-C1 strain could also degrade ZEN in Coicis Semen. If the initial content of ZEN in Coicis Semen was reduced from 90 μg·g~(-1) to 40.68 μg·g~(-1), the degradation rate could reach 54.80%. This study is expected to provide a new strain and application technology for the biological detoxification of ZEN in food processing products and Chinese medicinal materials.

Isolation and purification of neuraminidase from influenza vaccine and reference substance preparation / 中华微生物学和免疫学杂志

Objective:To establish a method for isolation and purification of neuraminidase from influenza vaccine and to prepare reference substance for quantitative detection of neuraminidase.Methods:Functional magnetic particles with specific affinity for neuraminidase were prepared. The method for separation and purification of neuraminidase was established based on the magnetic particles. The separation and purification conditions were optimized. The purity of neuraminidase was analyzed and the specificity was verified. The enzyme activity was determined and the protein was quantified.Results:The functional magnetic particles modified with 4-aminophenanthroline were successfully prepared and the method for isolation and purification of neuraminidase based on the magnetic particles was established. The purity of neuraminidase was 98.7%. The concentrations of neuraminidase isolated and purified from the monovalent stock solution of H1N1, H3N2, B/Victoria and B/Yamagate vaccines were 71.50, 100.58, 64.11 and 37.68 μg/ml, respectively, and the enzyme activity remained.Conclusions:The method for isolation and purification of influenza virus neuraminidase was established and the corresponding reference substance was prepared.

Immunogenicity of MDCK cell-based quadrivalent influenza split vaccine with different adjuvants in mice / 中华微生物学和免疫学杂志

Objective:To evaluate the immunogenicity of Madin-Darby canine kidney (MDCK) cell-based quadrivalent influenza split vaccine (MDCK-Va) combined with different adjuvants.Methods:Different doses of MDCK-Va and chicken embryo-based quadrivalent influenza split vaccine (egg-Va) were intramuscularly immunized BALB/c mice twice with an interval of three weeks. Serum samples were collected to detect antibody titers using hemagglutination inhibition (HI) assay. BALB/c mice were immunized with different doses of MDCK-Va combined with QS21, AddVax, PolyI∶C, CpG ODN 1826 and AddVax/PolyI∶C (Add/Poly), respectively. HI and microneutralization assays were used to detect antibody titers 21 d after the first and booster immunization. Spleen tissues were collected from the mice immunized with 10 μg MDCK-Va combined with the above adjuvants 5 d after the booster immunization to analyze spleen index and the types of spleen cells.Results:The immunoprotective effect of MDCK-Va was not inferior to that of egg-Va. MDCK-Va combined with each of the above adjuvants could induce higher HI antibody titer than MDCK-Va alone, especially the QS21/Va and Add/Poly/Va groups, and the differences were statistically significant. For H1N1 vaccine, the Pearson′s correlation coefficient ( r) between HI antibody and neutralizing antibody was 0.737-0.910, and for H3N2 subtype vaccine, the value of r was 0.839-0.947. Compared with the MDCK-Va group, the QS21/Va group showed significantly increased spleen index and decreased proportion of single lymphocytes. QS21 and Add/Poly were much better than other adjuvants in stimulating mouse splenic neutrophils and CD4/CD8 cells. Conclusions:Add/Poly had a stronger immune enhancement effect on MDCK-Va, suggesting that it was a potential adjuvant for MDCK-Va. The antibody titer detected by HI and MN assays had a strong positive correlation.

Treatment of open fracture of lower limb in high altitude area / 中国骨伤

OBJECTIVE@#To explore the treatment methods and experience of open fracture of lower limb in high altitude area.@*METHODS@#From January 2016 to January 2021, 62 patients with open fractures of lower limbs were treated by staged surgery with the concept of injury control orthopedics, emphasizing wound treatment and combining various fracture fixation methods. There were 51 males and 11 females, ranging in age from 14 to 59 years old, with a mean of (37.2±12.3) years old; and the course of disease ranged from 7 to 59 days, with a mean of (23.7±15.5) days. According to Gustilo Anderson classification, there were 14 cases of typeⅠ, 24 cases of typeⅡ, 14 cases of typeⅢA, 8 cases of typeⅢB and 2 cases of typeⅢC. The fracture repair and wound healing were observed, and the clinical efficacy was evaluated by Johner-Wruhs evaluation standard.@*RESULTS@#Fifty-five patients were followed up, and the duration ranged from 4 to 36 months, with a mean of (14.7±8.5) months, and 7 cases were lost to follow-up. According to Johner-Wruhs evaluation criteria, 33 cases got an excellent result, 16 good, 4 poor and 2 bad. The wound healing was poor in 2 cases, partial necrosis of Achilles tendon in 1 case, nonunion of fracture in 1 case and delayed healing of fracture in 2 cases.@*CONCLUSION@#It is an effective method to treat the open fracture of lower extremity in high altitude area to pay attention to the management of soft tissue injury, the management of wound moisturizing, staged operation of fracture and full protection of blood supply at the fracture end. Paying attention to the treatment of soft tissue injury and the management of wound moisturizing, staged operation of fracture and full protection of blood supply at the fracture end are effective methods for the treatment of open fracture of lower limbs in high altitude areas.

Three-hour postoperative pancreatic enzymes levels for early prediction of pancreatitis following ERCP / 中华全科医师杂志

Objective:To investigate the predictive value of 3-hour post-endoscopic retrograde cholangiopancreatography (ERCP) serum amylase and lipase levels for postoperative pancreatitis (PEP).Methods:Clinical data of patients who underwent ERCP from June 2017 to December 2018 in our hospital were retrospectively analyzed. Risk factors of PEP were examined with univariate and multivariate analysis. Receiver operator characteristic (ROC) curve for 3-h postoperative serum amylase and lipase was generated and the optimal cut-off levels for PEP prediction were calculated.Results:A total of 491 patients were enrolled in this study, and 29 patients developed PEP. Compared with the uncomplicated group, female gender [75.9% (22/29) vs. 49.1% (227/462)], pancreatogram [65.5% (19/29) vs. 32.0% (148/462)], pancreatic duct cannulation [27.6% (8/29) vs. 11.7% (54/462)] and coronary heart disease [17.2% (5/29) vs. 4.5% (21/462)] accounted for a significant higher proportion in PEP group (χ 2=7.79, 13.63, 6.25, 6.42, all P<0.05), while logistic regression analysis revealed that only pancreatic duct cannulation was an independent risk factor ( OR=3.574, 95 %CI: 1.315-9.713, P<0.05). Areas under the ROC curve of serum amylase and lipase at 3 h were 0.897 (95 %CI: 0.842-0.951, P<0.01) and 0.915 (95 %CI: 0.881-0.948, P<0.01), respectively ( Z=0.550, P>0.05). For amylase, ideal sensitivity(93.1%) and specificity(93.3%) could be obtained as cut-off levels>1.0 and 3.0 times the upper limit reference (ULR), respectively. Correspondingly, as cut-off levels for lipase>2.5 and 6.0 times the ULR, the sensitivity was 93.1% and specificity was 91.3%, respectively. Conclusion:Three-hour postoperative serum amylase and lipase are both ideal indicators for predicting PEP with similar clinical value.

Rhynchophylline nanosuspensions sustained-release tablets: Preparation, optimization and release evaluation in vitro / 中草药

Objective: To prepare the rhynchophylline nanosuspensions and lyophilized powder, and study its sustained-release tablets. Methods: Rhynchophylline nanosuspensions were prepared by microprecipitation combined with high pressure homogenization method, and the particle size and zeta potential were determined. Scanning electron microscopy (SEM) was employed to observe the appearances of nanosuspensions. Nanosuspensions were prepared into lyophilized powder using lactose as freeze-dried protectors. HPMC (hydroxypropyl methyl cellulose) was used as hydrophilic matrix to prepare the sustained-release tablets. Single factor investigation and orthogonal experiments were employed to optimize the formulation of rhynchophylline nanosuspensions sustained-release tablets, and the model fitting was also been studied. Results: The particle size and zeta potential of rhynchophylline nanosuspensions were (153.7 ± 4.9) nm and (-18.54 ± 1.32) mV, respectively. The appearances of rhynchophylline nanosuspensions were spherical or nearly spherical. After orthogonal optimization, the cumulative release rate of rhynchophylline nanosuspensions sustained-release tablets was 92.53% in 12 h. The optimized formulation of hydrogel matrix sustained-release tablets was better accorded with Higuchi model: ln(1-Mt/M∞)=0.286 0 t1/2-0.069 0 (r=0.992 4). The drug release from hydrogel matrix sustained-release tablets were controlled by diffusion and degradation. Conclusion: The obtained rhynchophylline nanosuspensions has small particle size. The prepared hydrogel matrix sustained-release tablets can control the release of rhynchophylline nanosuspensions in a slow characteristic.

Effect of Hp infection on the inflammatory factors gastrointestinal hormones and insulin resistance in patients with type 2 diabetes mellitus / 国际检验医学杂志

Objective To investigate the effect of Hp infection on the inflammatory factors, gastrointestinal hormones and insulin resistance in patients with type 2diabetes mellitus.Methods A total of 86patients with type 2diabetes mellitus treated in a hospital from May 2014to October 2016were enrolled in the study.All patients underwent 13C urea breath test after admission, and according to the test results, patients were divided into the observation group, which was type 2diabetes mellitus with Hp infection (n=46) and the control group, which was type 2diabetes mellitus without Hp infection (n=40) .Inflammatory factors such as highsensitivity C reactive protein (hs-CRP) , serum interleukin 6 (IL-6) , tumor necrosis factorα (TNF-α) , gastrointestinal hormones such as serum gastrin (GAS) , somatostatin (SS) and fasting blood glucose (FPG) , fasting insulin (FINS) , insulin resistance index (HOMA-IR) were compared between the two groups, and the correlation of the observation indicators with Hp infection was explored.Results Levels of hs-CRP, IL-6, TNF-αand GAS, FPG, FINS and HOMA-IR in the observation group were significantly higher than those in the control group, while SS was significantly lower than that in the control group, the difference was statistically significant (P<0.05) .Further Sperman correlation analysis showed that hs-CRP, IL-6, TNF-α, GAS, FINS and HO-MA-IR were positively correlated with Hp infection (r=0.452, 0.350, 0.398, 0.389, 0.421, 0.568, P<0.05) , and SS was negatively correlated with Hp infection (r=-0.401, P<0.05) .Conclusion Hp infection is closely related to inflammatory reactions, gastrointestinal hormone disorders and insulin resistance in patients with type 2diabetes mellitus, and taking reasonable preventive measures is the key to delay the progression and deterioration of type 2diabetes mellitus.

Meta-analysis of the clinical effect and safety of early oral feeding in patients with acute pancreatitis / 国际外科学杂志

Objective To evaluate the clinical effect and safety of early oral feeding versus delayed feeding in patients with acute pancreatitis using Meta-analysis.Methods An electronic search of PubMed,Cochrane Library,Embase,Web of Science,CNKI,Wanfang database and VIP database were retrieved from their inception to March 2019 to identify relevant literatures,by taking'acute pancreatitis,oral,food intake,refeeding,feeding,diet'as the keywords for retrieval.The evaluation indexes included hospitalization time,incidence of abdominal pain after feeding,incidence of nausea and vomiting after feeding,incidence of hyperglycemia and leucocyte level after feeding.To select literatures and extractdata strictly according to inclusion and exclusion criteria,then conducted data analysis by Revman 5.3 provided by Cochrane collaboration.Results A total of 9 articles,involving 856 patients,were included with 425 in the early feeding group and 431 in the delayed feeding group.The length of hospitalization in early feeding group was shorter than that in delayed feeding group (MD =-2.37,95％ CI:-3.45--1.28,P ＜ 0.000 1).The incidence of abdominal pain (P =0.67),nausea and vomiting (P =0.77),hyperglycemia (P =0.54),and leukocyte level (P =0.26) were not significantly different between two groups.Conclusion Early oral feeding is safe and effective for patients with acute pancreatitis,which can significantly shorten the hospital stay without increasing the incidence of adverse reactions.

Study on matrix sustained release tablets of brucine solid lipid nanoparticles / 中草药

Objective To prepare brucine solid lipid nanoparticles (SLN) and its lyophilized powder, and then hydrogel matrix sustained-release tablets (HMST) of brucine SLN (SLN-HMST) were prepared. The factors that may influence drug release in vitro and release mechanism were also investigated in present study. Methods Based on single factor test, orthogonal test was designed to gain the optimum prescription. Zero-order, First-order and Higuchi models were used for the model fitting of drug release. Ritger-Pappas models were employed to study release mechanism of brucine SLN-HMST. Results Brucine SLN-HMST was better agreed with First-order kinetics model. The equation was ln(1-Mt/M∞) = -0.212 1 t + 0.106 4 (r = 0.992 3). The cumulative release could achieve 91.48% in 12 h. The sustained release features were obviously. The drug release from the tablets was controlled by diffusion and degradation of the matrix. Conclusion The prepared brucine SLN-HMST can deliver drug continually for 12 h with good reproducibility.

Two kinds of luteolin solid dispersion: Preparation, characterization, and pharmacokinetic behaviors in SD rats in vivo / 中草药

Objective To prepare luteolin solid dispersions (Lut-SD) and luteolin phospholipids complex solid dispersions (Lut-PC-SD), and compare the effects of two kinds of solid dispersions on the bioavailability in vivo. Methods PVP K30 was employed as carrier, and solvent evaporation method was used to prepare Lut-SD and Lut-PC-SD. Their existential state of luteolin in solid dispersions was analyzed by X-ray power diffraction (XRPD). The solubility and dissolution rate were also studied. SD rats in each group were administered intragastrically with Lut, Lut-SD, and Lut-PC-SD, respectively. Their blood samples were collected at different time intervals. Diosmetin was used as internal standard, the concentration of Lut in blood was analyzed by HPLC, and the main pharmacokinetic parameters were obtained. Results The results of XRPD indicated that Lut showed an amorphous state in Lut-SD and Lut-PC-SD. The solubility of Lut was enhanced from (61.09 ± 0.09) μg/mL to (365.33 ± 0.38) μg/mL and (401.14 ± 0.19) μg/mL by Lut-SD and Lut-PC-SD, repectively. The dissolution of Lut was also improved greatly by the two kinds of solid dispersions. Compared to Lut, the bioavailability of Lut-SD and Lut-PC-SD was enhanced to 150.10% and 204.52%, repectively. Conclusion Lut-SD and Lut-PC-SD both could enhance the bioavailability of Lut in SD rats notably. In addition, Lut-PC-SD could give a better effect.

Aseptic loosening of artificial joint:old story and advances / 医学研究生学报

Aseptic loosening is the most common chronic complication of total joint arthroplasty,which significantly shortens the life span of prosthesis and leads to mental stress and heavy economic burden of patients.Hence,how to overcome aseptic loosening is a focus in the research of joint arthroplasty.A series of mechanisms participates the pathogenesis of aseptic loosening,including oste-olysis induced by wear particles,micromotion,stress shielding,high fluid pressure,endotoxin and genetic factors.Osteolysis induced by wear particles is the most recognized mechanism.The latest study suggested that autophagy,epigenetic regulation and immunologic derangement all participate the development of osteolysis induced by wear particles.However,the prevention,treatment and even the concept of aseptic loosening is still controverial.We hope that the future study will provide a definite mechanism for aseptic loosening and provide new theoretic evidence for the prevention and treatment of aseptic loosening.

Mechanism of Chronic Stress-induced Reduced Atherosclerotic Medial Area and Increased Plaque Instability in Rabbit Models of Chronic Stress / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Chronic stress contributes to increased risks of atherosclerotic diseases including heart disease, stroke, and transient ischemic attack. However, its underline mechanisms are poorly understood. This study aimed to elucidate the mechanism via which chronic stress exerts its effect on atherosclerosis (AS).</p><p><b>METHODS</b>Fifty male New Zealand white rabbits were used. Aortic balloon-injury model was applied. Both social stress and physical stress methods were adopted to establish chronic stress models. The lumen stenotic degree, intimal and medial areas, maximum fibrous cap thickness, and plaque contents were measured with histological sections. Proteomic methods were applied to detect protein changes in abdominal aortas to identify the specialized mediators. Real-time reverse transcription-polymerase chain reaction was used for further verification and investigation.</p><p><b>RESULTS</b>The stress rabbits exhibited lower body weight, worse fur state, more inactivity behavior, and higher serum cortisol level. Chronic stress was significantly associated with the decreased medial area and increased plaque instability, which was manifested by thinner fibrous caps, larger lipid cores, more macrophages, and new vessels but fewer smooth muscle cells and elastic fibers. After chronic stress, the apoptosis-related genes UBE2K, BAX, FAS, Caspase 3, Caspase 9, and P53 were upregulated, and BCL-2/BAX was down-regulated; the angiogenesis-related genes ANG and VEGF-A were also highly expressed in atherosclerotic arteries.</p><p><b>CONCLUSIONS</b>Rabbit models of chronic stress were successfully established by applying both social stress and physical stress for 8 weeks. Chronic stress can reduce AS tunica media and accelerate plaque instability by promoting apoptosis and neovascularization.</p>

Etiologic diagnosis of small bowel ulcer with double balloon enteroscopy / 中华全科医师杂志

Objective To assess the value of double balloon enteroscopy (DBE) in etiological diagnosis of small bowel ulcer.Methods The clinical and endoscopic data of patients undergoing DBE at the Department of Gastroenterology, Renmin Hospital of Wuhan University from January 2010 to January 2016 were reviewed.For patients with small bowel ulcer revealed by DBE, the etiologic diagnosis was retrospectively analyzed.Results One hundred and three patients with small bowel ulcer were included in the analysis.The initial diagnosis based on morphology of ulcer was Crohn's disease (58.3%, 60/103), intestinal tuberculosis (18.4%,19/103), etiology unknown (24.3%, 25/103).The final diagnosis comprehensively based on pathology, clinical data and disease outcome was Crohn's disease (44.7%,46/103), intestinal tuberculosis (18.4%,19/103), lymphoma (3.8%,4/103), eosinophils enteritis (1.9%,2/103), non-steroidal anti-inflammatory drug enteritis (1.9%,2/103), adenocarcinoma (1.0%,1/103), stroma tumor (1.0%,1/103),ischemic enteritis (1.0%,1/103), vascular malformation (1.0%,1/103), Henoch-Schonlein purpura (1.0%,1/103), etiology unknown (24.3,25/103).The accurate diagnosis rate and misdiagnosis rate for Crohn's disease by DBE were 73.3%(44/60)and 10.0%(6/60).The accurate diagnosis rate and misdiagnosis rate for intestinal tuberculosis by DBE were 14/18 and 2/18.Conclusion There are diversity and complexity in etiologic diagnosis of small bowel ulcer.The diagnosis should be based on the comprehensive analysis of morphology of ulcer, endoscopic and surgical pathology and clinical data.Follow up of small bowel ulcer by DBE may provide important information for etiologic diagnosis.

The diagnosis and treatment of obscure gastrointestinal bleeding using total enteroscopy with double balloon enteroscopy / 中华消化内镜杂志

Objective To investigate the feasibility and application value of total enteroscopy with double balloon enteroscopy(DBE)for diagnosis and treatment of patients with obscure gastrointestinal bleeding(OGIB). Methods The clinical and endoscopic data of patients underwent DBE for OGIB in the Department of Gastroenterology,Remin Hospital of Wuhan University from January 2010 to December 2015 were retrospectively analyzed. Results Total enteroscopy was indicated in 36.3%(136/375)of patients. The success rate was 86.0%(117/136)and complication rate was 1.5%(2/136). Negative findings,non-small bowel lesions and small bowel lesions were detected in 44.4%(52/117), 6.8%(8/117), and 48.7%(57/117)of patients with total enteroscopy. Re-bleeding occurred in 8.9%(4/45)of patients with negative total enteroscopy,while 1 small bowel mesenchymoma and 1 gastric fundus Dieulafoy′s lesion were revealed subsequently. Re-bleeding occurred in 33.3%(5/15)of patients with incomplete enteroscopy,and 1 small bowel polyp and 1 small bowel angiectasis were revealed subsequently. Conclusion DBE can complete total enteroscopy within one day and provide important clinical information of OGIB. Non-small bowel lesions,small bowel lesion missed by DBE and potential bleeding lesions in small bowel beyond the reach of DBE should be considered in patients with negative enteroscopy.

The correlation of serum SOD and CD4+CD25+regulatory T cell in the mechanism of severe sepsis / 实用医学杂志

Objective To evaluate the condition of oxidative stress and immunosuppression in early stage of severe sepsis,and investigate the correlation between them. Methods A prospective random control study in-cluded patients group(n=51)and control group(n=31). The concentration of serum superoxide dismutase was measured by enzyme linked immunosorbent assay(ELISA),CD4+CD25+Treg% was measured by flow cytometry , respectively. The difference between two groups was compared and the correlation between parameters in patients group was evaluated. Results The concentration of serum SOD was lower than control group (P < 0.01). CD4+CD25+Treg% significantly high,compared to the control group(P < 0.01). There was no strong correlation be-tween parameters in patients group. Conclusion Oxidative stress and immunosuppression are exist in the early stage of severe sepsis.

Mineral Intake in Urban Pregnant Women from Base Diet, Fortified Foods, and Food Supplements: Focus on Calcium, Iron, and Zinc / 生物医学与环境科学（英文）

In the Chinese national nutrition surveys, fortified foods were not investigated separately from the base diet, and the contribution of fortified foods to micronutrients intake is not very clear. This study investigated the diet, including fortified foods and food supplements, of urban pregnant women and analyzed the intake of calcium, iron, and zinc to assess the corresponding contributions of fortified foods, food supplements, and the base diet. The results demonstrated that the base diet was the major source of calcium, iron, and zinc, and was recommended to be the first choice for micronutrients intake. Furthermore, consumption of fortified foods and food supplements offered effective approaches to improve the dietary intake of calcium, iron, and zinc in Chinese urban pregnant women.