Screening specific minimum amino acid sequence triggering immunity to enterovirus 71 / 中华微生物学和免疫学杂志

Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.

Effect of posterior tibial slope on the short-term outcome in mobile-bearing unicompartmental knee arthroplasty / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the relationship between postoperative knee function and the sagittal position of tibial component in unicompartmental knee arthroplasty (UKA).@*METHODS@#We retrospectively enrolled the patients who underwent UKA from January 2016 to May 2020. They were assigned into 2 groups according to postoperative posterior tibial slope (PTS): the normal PTS group (PTS≥3° and PTS < 8°) and the abnormal PTS group (PTS < 3° or ≥8°). The patients were followed up for at least 12 months. The postoperative Knee Society Clinical Score (KSS-C), Knee Society Functional Score (KSS-F) and knee range of motion (ROM) were compared between the two groups.@*RESULTS@#A total of 72 patients (82 knees) were included with 51 patients (58 knees) in PTS normal group and 21 patients (24 knees) in PTS abnormal group. All the patients were followed up with median of 23.6 months. There was no significant difference in the general data [gender, age, body mass index (BMI)], pre-operative knee range of motion, preoperative KSS-C score and KSS-F score (P > 0.01). The KSS-C score, KSS-F score, and knee range of motion significantly improved after surgery (P < 0.01) for all the patients. The postoperative KSS-C score in normal PTS group (88.76±2.79) was significantly higher than the KSS-C score in abnormal PTS group (84.42±3.35, P < 0.01), but no significant difference between the 2 groups was observed in postoperative KSS-F score and knee range of motion (P > 0.01). In addition, there was no correlation between the change of PTS and postoperative KSS-C score (r=-0.034, 95%CI: -0.247 to 0.186, P = 0.759), KSS-F score (r = -0.014, 95%CI: -0.238 to 0.198, P = 0.901) and knee range of motion (r= 0.045, 95%CI: -0.214 to 0.302, P = 0.686).@*CONCLUSION@#The posterior tibial slope between 3° and < 8° can be recommended to improve knee joint function in mobile UKA, and excessive or insufficient PTS should be avoided.

Identification of Stripe Rust Resistance Genes in Common Wheat Cultivars From the Huang-Huai-Hai Region of China.

Wheat stripe (yellow) rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a serious fungal disease worldwide, especially in the Huang-Huai-Hai region, a main wheat production area in China. Gene postulation, molecular testing, and pedigree analysis were conducted to determine the presence of stripe rust resistance genes to 15 Pst races in 66 selected commercial wheat cultivars released from 2000 to 2016. In addition, races CYR32, CYR33, and CYR34 were used to evaluate resistance to Pst at the adult-plant stage of wheat in the field. Four Yr genes (Yr9, Yr10, Yr26, and Yr32) were postulated in 24 wheat cultivars either singly or in combination. Thirty-six cultivars might contain unknown Yr genes, whereas no identified Yr gene was postulated in six cultivars. Yr9 was detected at a frequency of 28.8%, and no cultivars carried Yr5, Yr15, or Yr18. Ten cultivars (15.2%) exhibited adult-plant resistance in the field tests with three predominant races. Three cultivars (Langyan 43, Xinong 889, and Yunfeng 139) had all-stage resistance. These results are useful to growers selecting cultivars and to breeders aiming to use more resistance genes to develop new cultivars with effective resistance in order to reduce stripe rust damage.

[The protective effects of lycium barbarum polysaccharides on retinal neurons in diabetic rats and its mechanism].

OBJECTIVE: To clarify whether lycium barbarum polysaccharides (LBP) have protective effects on retina neuronal cells in diabetic rats and to identify the related mechanism involved in this process. METHODS: Eighteen SD rats were randomly divided into 3 groups ( n= 6): normal control group (NC), diabetes mellitus group (DM) and LBP-treatment group (DM+LBP). The diabetic rat model was induced by single intraperitoneal injection of streptozotocin (STZ). The rats in DM+LBP group were treated with LBP at the dose of 1 mg/kg by gavage, once a day for 12 weeks. After the treatment, the weight and blood glucose, the generation of reactive oxygen species (ROS), the surviving retinal ganglion cells (RGCs) and amacrine cells and the protein expressions of nuclear factor E2-related factor 2 (Nrf2) and the heme oxygenase-1 (HO-1) were detected. RESULTS: The successful rate of diabetic model was 100%. Compared with NC group, the rats of DM group caused weight loss, elevated blood glucose, a marked increase of ROS generation and a significant decrease in the number of RGCs and amacrine cells (P＜0.01), and these effects were diminished or abolished by LBP treatment. Meanwhile, LBP significantly increased the expressions of Nrf2 and HO-1 in the retinas of diabetic rats (P＜0.01). CONCLUSION: LBP can improve retinal oxidative stress and exert beneficial neuroprotective effects in diabetic rats, and its mechanism may be associated with the activation of the Nrf2/HO-1 antioxidant pathway.

Effects of live attenuated polio vaccine on the immunogenicity of live attenuated rotavirus vaccine / 中华微生物学和免疫学杂志

Objective To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus ( RV) vaccine. Methods Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups:interval vaccination group ( Rotarix and OPV were vaccinated on different days) and simultane-ous vaccination group ( Rotarix and OPV were vaccinated on the same day) . Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups. Results The seroconversion rate of serum RV-IgA in month 2 was 73. 84％ in the interval vaccination and 63. 95％ in the simultaneous vaccination group, and the difference between them was statistically significant (P<0. 05). The geometric mean concen-trations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0. 05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0. 05). Conclusions Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenua-ted rotavirus vaccine, especially the maintenance of RV-IgA antibody level.

The protective effects of lycium barbarum polysaccharides on retinal neurons in diabetic rats and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To clarify whether lycium barbarum polysaccharides (LBP) have protective effects on retina neuronal cells in diabetic rats and to identify the related mechanism involved in this process.@*METHODS@#Eighteen SD rats were randomly divided into 3 groups ( n= 6): normal control group (NC), diabetes mellitus group (DM) and LBP-treatment group (DM+LBP). The diabetic rat model was induced by single intraperitoneal injection of streptozotocin (STZ). The rats in DM+LBP group were treated with LBP at the dose of 1 mg/kg by gavage, once a day for 12 weeks. After the treatment, the weight and blood glucose, the generation of reactive oxygen species (ROS), the surviving retinal ganglion cells (RGCs) and amacrine cells and the protein expressions of nuclear factor E2-related factor 2 (Nrf2) and the heme oxygenase-1 (HO-1) were detected.@*RESULTS@#The successful rate of diabetic model was 100%. Compared with NC group, the rats of DM group caused weight loss, elevated blood glucose, a marked increase of ROS generation and a significant decrease in the number of RGCs and amacrine cells (P＜0.01), and these effects were diminished or abolished by LBP treatment. Meanwhile, LBP significantly increased the expressions of Nrf2 and HO-1 in the retinas of diabetic rats (P＜0.01).@*CONCLUSION@#LBP can improve retinal oxidative stress and exert beneficial neuroprotective effects in diabetic rats, and its mechanism may be associated with the activation of the Nrf2/HO-1 antioxidant pathway.

Effects of live attenuated polio vaccine on the immunogenicity of live attenuated rotavirus vaccine / 中华微生物学和免疫学杂志

Objective@#To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus (RV) vaccine.@*Methods@#Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups: interval vaccination group (Rotarix and OPV were vaccinated on different days) and simultaneous vaccination group (Rotarix and OPV were vaccinated on the same day). Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups.@*Results@#The seroconversion rate of serum RV-IgA in month 2 was 73.84% in the interval vaccination and 63.95% in the simultaneous vaccination group, and the difference between them was statistically significant (P<0.05). The geometric mean concentrations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0.05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0.05).@*Conclusions@#Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenuated rotavirus vaccine, especially the maintenance of RV-IgA antibody level.

Validation of a cell infection-based quantitative RT-PCR for evaluation of rotavirus vaccine potency / 中华微生物学和免疫学杂志

Objective To validate a cell infection-based quantitative RT-PCR for evaluating the potency of rotavirus vaccine. Methods According to the ICH ( the International Council for Harmonization) Harmonised Tripartite Guideline, the method was validated for its specificity, accuracy, precision, linearity and robustness. Results The method had good specificity as it could only amplify and detect the corre-sponding type of rotavirus strain. The recovery rates for determining the potency against rotaviruses of G2, G3 and G4 types were 97％ to 108％. The percent coefficient of variation ( CV) of both intra-plate and in-ter-plate precision was≤2. 62％, while the intraday and interday CV was≤1. 76％ and≤2. 27％, respec-tively. The CV between the two experimenters was≤7. 68％. The linearity range of the method was 4. 4-6. 5 UI for G2 type rotavirus, 3. 9-8. 3 UI for G3 type and 3. 5-8. 1 UI for G4 type. Good robustness was observed using the cells of 140 to 160 generations. Conclusions The cell infection-based quantitative RT-PCR was shown to have satisfactory specificity, accuracy, precision, linearity and robustness, suggesting that it was a suitable method for evaluating the potency of multivalent rotavirus live vaccines.

Should kidney volume be used as an indicator of surgical occasion for patients with autosomal dominant polycystic kidney disease?

To investigate the best surgical occasion of laparoscopic cyst decortications (LCDs) in patients with autosomal dominant polycystic kidney disease (ADPKD), in accordance with the renal volume (RV). We retrospectively analyzed 135 (65 male and 70 female) patients with ADPKD who underwent LCD between June 2011 and October 2015. Patients were divided into 4 groups according to the volume of the operated kidney measured from computed tomography scans: group A (28 patients, RV < 500 mL), group B (63 patients, RV = 500-1000 mL), group C (30 patients, RV = 1000-1500 mL), and group D (14 patients, RV > 1500 mL). We studied postoperative indicators at least 1-year follow-up. For each RV group, therapeutic responses of LCD in these patients with ADPKD were assessed by improvement of clinical parameters and manifestations. A significant glomerular filtration rate (GFR) improvement was found in RV group B (31.8 ±â€Š11.1 mL/min; final GFR 36.9 ±â€Š12.7 mL/min; P < 0.01), and RV group C (21.1 ±â€Š8.7 mL/min; final GFR 27.4 ±â€Š9.2 mL/min; P < 0.01). RV group C had much higher GFR improvements than did RV group B (P < 0.01). In addition, refractory pain in patients of RV groups B, C, and D was much relieved by LCD treatment. Compared with other RV groups, blood pressures in patients with ADPKD of RV group D were also improved (P < 0.01). Our study indicates that RV could be used to evaluate LCD clinical outcomes in patients with ADPKD. The results of LCD for patients with ADPKD with RV between 500 and 1500 mL were encouraging, especially with regards to renal function improvement and pain relief. Therefore, RV may become a useful marker to predict the timing of LCD surgery in patients with ADPKD.

High Relative Parasitic Fitness of G22 Derivatives is Associated with the Epidemic Potential of Wheat Stripe Rust in China.

In total, 13 commercial wheat cultivars around China and four races of Puccinia striiformis f. sp. tritici (namely, CYR32, CYR33, G22-9, and G22-14) were employed for a test of relative parasitic fitness (RPF) using the drop method. The RPF values were measured, including the urediniospore germination rate, the latent period, the uredinial length, the uredinial density, the infection area, the sporulation intensity, the lesion expansion speed, and the sporulation period. The results indicated that the parameters of relative parasitic fitness of the four P. striiformis f. sp. tritici races on the 13 wheat cultivars were significantly different (P = 0.00) in sporulation intensity, lesion expansion speed, uredinial length, sporulation period, uredinial density, and latent period. The urediniospore germination rates of the four P. striiformis f. sp. tritici races for the test were significantly different (P = 0.00), whereas no correlation with the different cultivars was observed (P = 1.00). The infection areas of the tested races on the different cultivars were significantly different (P = 0.00) but there were no obvious manifestations among the various races (P = 0.20). Principal component analysis (PCA) showed that the sporulation intensity represented sporulation capacity and scalability, the latent period indicated infection ability, and the urediniospore germination rate represented urediniospore vigor, all of which fully contributed to the RPF in the interaction of the four races and 13 wheat cultivars, which was calculated by the following formula: RPF = (sporulation intensity × urediniospore germination rate)/latent period. The sporulation and infection of G22-9 on the 13 large-scale cultivated cultivars were the highest, and the RPF of G22-9 was higher than that of the predominant races, CYR32 and CYR33. This result suggested that G22-9 could become a new predominant race and potentially cause epidemics of wheat stripe rust in China. To prevent potential epidemics, susceptible wheat cultivars should be withdrawn from production and breeding programs should reduce the use of Yr10 and Yr26 and use other more effective resistance genes in combination with nonrace-specific resistance for developing wheat cultivars with durable resistance to stripe rust.

Progress in developing Norovirus vaccines based on virus-like particles / 中华微生物学和免疫学杂志

Human norovirus (NoV) is the major cause of human acute non-bacterial gastroenteritis worldwide. The development of NoV vaccines is limited by the inability to culture cells in vitro and the lack of small animal models. Thus, traditional methods used to prepare live attenuated vaccines are not suitable for preparing NoV vaccines. Subunit vaccines against NoV infection, especially virus-like particle (VLP)-based vaccines, have outstanding advantages in the research and development of NoV vaccines. In this review, we focused on reviewing recent advances in the fields of NoV VLP-based vaccines and the development of NoV VLP-based vaccines using different expression systems as well as identifying the research direction for NoV VLP-based vaccines.

Hierarchical Porous Intercalation-Type V2 O3 as High-Performance Anode Materials for Li-Ion Batteries.

As intercalation-type anode materials for Li-ion batteries (LIBs), the commercially used graphite and Li4 Ti5 O12 exhibit good cycling and rate properties, but their theoretical specific capacities are too low to meet the ever-growing demands of high-energy applications such as electric vehicles. Therefore, the development of new intercalation-type anode materials with larger capacity is very desirable. Herein, we design and synthesize novel 3 D hierarchical porous V2 O3 @C micro/nanostructures consisting of crumpled nanosheets, through self-reduction under annealing from the structurally similar VO2 (B)@C precursors without the addition of any other reducing reagent or gas. Excitingly, it is found for the first time through ex situ XRD technology that V2 O3 is a new, promising intercalation-type anode material for LIBs with a high capacity. V2 O3 @C micro/nanostructures can deliver a large capacity of 732 mAh g-1 without capacity loss at 100 mA g-1 even after 136 cycles, as well as exhibiting excellent cycling and rate performances. The application of V2 O3 for Na-ion batteries (NIBs) is elaborated for the first time, and excitingly, it is found that V2 O3 @C micro/nanostructures may be promising anode materials for NIBs.

Induction chemotherapy followed by concurrent chemoradiation and nimotuzumab for locoregionally advanced nasopharyngeal carcinoma: preliminary results from a phase II clinical trial.

Overexpression of epidermal growth factor receptor can be found in more than 80% of patients with locoregionally advanced nasopharyngeal carcinoma and is associated with shorter survival. In this work, we evaluated the feasibility of adding nimotuzumab to chemoradiation in locoregionally advanced nasopharyngeal carcinoma. Twenty-three patients with clinically staged T3-4 or any node-positive disease were enrolled. They were scheduled to receive one cycle of induction chemotherapy followed by intensity-modulated radiotherapy, weekly administration of nimotuzumab and concurrent chemotherapy. Results showed that all patients received a full course of radiotherapy, 19(82.6%)patients completed the scheduled neoadjuvant and concurrent chemotherapy, and 22(95.7%) patients received ≥6 weeks of nimotuzumab. During the period of concurrent chemoradiation and nimotuzumab, grade 3-4 toxicities occurred in 14(60.9%) patients: 8 (34.8%) had grade 3-4 oral mucositis, 6(26.1%) had grade 3 neutropenia, and 1(4.3%) had grade 3 dermatitis. No acne-like rash was observed. With a median follow-up of 24.1 months, the 2-year progression-free survival and overall survival were 83.5% and 95.0%, respectively. In conclusion, concurrent administration of chemoradiation and nimotuzumab was well-tolerated with good compliance. Preliminary clinical outcome data appear encouraging with favorable normal tissue toxicity results comparing with historical data of concurrent chemoradiation plus cetuximab.

[Application of 3D digital orthopedic techniques in treatment of acetabular fracture].

OBJECTIVE: To evaluate the therapeutic effect of three-dimensional digital orthopedic techniques in treatment of acetabular fractures. METHODS: We retrospectively analyzed 50 cases of acetabular fracture treated between March, 2007 and December, 2013. The lamellar CT scanning data were imported into Mimics software, and 3D anatomical models of the pelvic and proximal femur were reconstructed. Computer-assisted analysis was carried out to understand the condition of fractures and simulate fracture reduction. The pelvic models were manufactured by rapid prototyping technique for definite diagnosis and typing of acetabular fractures and subsequent surgical treatment. RESULTS: Three-dimensional reconstruction images and rapid prototyping pelvic models faithfully represented the findings in operations. Preoperative simulation of the operation shortened the time of operation and reduced the volume of bleeding in the operation. All the patients were followed up for 6 to 24 months. According to Matta imaging score, anatomical reduction was achieved in 41 cases and satisfactory reduction in 9 cases. According to the Harris functional criteria, 32 patients had excellent, 12 had good and 6 had acceptable outcomes with a rate of excellent and good outcomes of 88%. CONCLUSION: Three-dimensional digital orthopedic techniques allow accurate display of the acetabulum and the spatial relation of the anatomic structures to assist in fracture diagnosis, typing and treatment.

Application of TG-ROC analytical method in detection reagent quality evaluation / 国际检验医学杂志

Objective To compare the relationship between the enzyme‐linked immunosorbent assay(ELISA) reagent and West‐ern blot(WB) confirmation reagent for analyzing the quality lever of human T‐cell lymphotropic virus(HTLV) detection reagent . Methods The WB confirmation reagent was used to detect anti‐HTLV antibody in 156 human serum samples of ELISA prelimina‐ry screening positive .The ELISA cut‐off value(optimal value) was selected by using the two‐graph receiver operating characteristics (TG‐ROC) analytical method .The two‐by‐two table analysis was constructed to analyze the consistency of results detected by the two methods ,moreover the McNemar test was used to evaluate the consistency of detection results .The quality level of HTLV de‐tection reagent was comprehensively evaluated .Results Among 156 serum samples of ELISA preliminary screening positive ,only 40 samples were positive by the WB confirmation ,and other 116 samples were negative .The sensitivity and specificity of ELISA de‐tection reagent obtained by TG‐ROC analysis were 97 .5% and 45 .7% respectively ,the TG‐ROC test also indicated that the detec‐tion results had significant difference between ELISA and WB(P<0 .05) .By adjusting the cut‐off value ,the sensitivity and specific‐ity of ELISA were increased to 88 .8% (parametric method) .In the comparison of the parametric method and the non‐parametric method ,the obtained areas under the curve(AUC) was 0 .923 5(parametric method) ,their results were basically consistent .Conclu‐sion Although above results indicate that the detection results of ELISA reagent are different from those of WB ,but adjusting the cut off value can increase its sensitivity and specificity ,thus increases the reliability of diagnosis result .

Norovirus P particle and its application / 中华微生物学和免疫学杂志

The challenges posed by norovirus infections to global health are increasing accompa-nied by the rapid rate of the genetic and antigenic evolution of circulating noroviruses. Due to lack of in vitro culture cells and small animal models, norovirus vaccine cannot be prepared by using traditional techniques. With the in-depth understanding and study of norovirus, the subunit vaccines against norovirus infection based on P particles have been developed and presented the characteristics of easily expressed, low cost, high immunogenicity, stable structure and so on. In addition, norovirus P particle has been used as a subvi-ral nanoparticle for vaccine development against other viruses and for antibody production against chronic dis-ease ( Alzheimer′s disease) , which benefits from the accommodation of foreign antigens in the three loops of P particle. In this review, we describe the progresses in the field of P particle related vaccines for providing suggestions about the research and development of multivalent vaccines in China.

Preparation and investigation of hydrolyzed polyacrylonitrile as a preliminary biomedical hydrogel.

BACKGROUND: Hydrolyzed polyacrylonitrile (HPAN) has attracted much attention as a hydrogel for a broad range of biomedical applications. Therefore, in this study, we prepared HPAN derivatives with controllable compositions by the radical polymerization of acrylonitrile (AN), methacrylic acid (MAA) and N-isopropylacrylamide (NIPAM) monomers. RESULTS: The prepared poly(AN-co-MAA-co-NIPAM) copolymers had different ratios of AN, MAA, and NIPAM and molecular weights ranging from 2000 to 50,000. The copolymers were prepared as films to examine their properties. The prepared copolymer films showed different solubilities, contact angles, and swelling ratios. The properties of the copolymer films were affected by the hydrophobic PAN segments and the hydrophilic PMAA or PNIPAM segments. CONCLUSION: Thus, we conclude that introducing PMAA and PNIPAM segments with different ratios and lengths into PAN segments could represent a method of controlling the hydrogel properties of copolymers.

The Effects of ASMase Mediated Endothelial Cell Apoptosis in Multiple Hypofractionated Irradiations in CT26 Tumor Bearing Mice.

BACKGROUND: To investigate the effects of ASMase mediated endothelial cell apoptosis in multiple hypofractionated irradiations in CT26 tumor bearing mice. MATERIALS AND METHODS: Thirty-five CT26 tumor bearing mice were subjected to single ionizing radiation (IR) of 0, 3, 6, 9, 12, 15, 18 Gy. Eight hours after IR, the mice were sacrificed and tumor tissues were used for CD31 immunohistochemistry staining, TUNEL and CD31 double staining, ASMase activity assay. Then 6 and 12 Gy were chosen for multiple hypofractionated IR experiments according to the above results. Each time after IR, 5 mice were sacrificed and assayed as above. RESULTS: The ASMase activities were increased significantly after a single IR of 12 Gy or higher which was accompanied with remarkable increased endothelial cell apoptosis and decreased MVD. For 6 Gy which was not high enough to trigger ASMase activation, after 2 or more times of IR, the ASMase activities were significantly increased accompanied with remarkable increased endothelial cell apoptosis and decreased MVD. While for 12 Gy, after 2 or more times of IR, the ASMase activities and endothelial cell apoptosis rates were maintained without remarkable increase; however, the MVD was significantly decreased. What's more, the cancer cell apoptosis rates were significantly increased after multiple IR for both 6 Gy and 12 Gy. CONCLUSIONS: ASMase mediated endothelial cell apoptosis may play an important role in the process of multiple hypofractionated IR for CT26 colorectal carcinoma.

The effects of lymph after intestinal ischemia/reperfusion on the inflammatory factors and Toll-like receptor 4 ligand high mobility group box-1 in Toll-like receptor 4 deficient mice / 中华临床营养杂志

Objective To investigate the effects of lymph from ischemic/reperfused intestine on the inflammatory factors and Toll-like receptor 4 (TLR4) ligand high mobility group box-1 (HMGB1) in TLR4 deficient (TLR4-/-) mice.Methods A total of 20 SD rats weighing (300 ±20) g were randomly assigned into two groups:lymph drainage group (group N,lymph drainage for 180 minutes without other treatment) and intestinal ischemia/reperfusion group (group I/R,draining the lymph for 180 minutes while clipping the superiormesenteric artery for 60 minutes followed by 120-minute reperfusion).Thirty-two TLR4-/-mice and thirty-two C57BL/6 wild type (WT) mice were each divided into 4 sub-groups (n =8),injected with different fluids through the caudal vein:group N with normal lymph;group I/R with I/R lymph;group Edt with endotoxin;group HMGB1 with HMGB1 protein.The mice were sacrificed 180 minutes after the injection for sample collection.Results The levels of endotoxin and HMGB1 in the lymph drainage of the group I/R rats were significantly higher than that of the group N rats [(0.034 ± 0.050) Eu/ml vs.(0.017 ± 0.023) Eu/ml,P =0.033;(4.293 ± 0.883) ng/ml vs.(0.509 ± 0.128) ng/ml,P =0.006].In the mice injected with HMGB1,the mucosa thickness and villus height in the ileum of the WT mice were significantly lower than that of the TLR4-/-mice [(335.8±43.2) μmvs.(602.1±37.5) μm,P=0.000;(273.0±31.7) μm vs.(404.5 ± 18.6) μm,P =0.000];in both WT and TLR4-/-mice injected with the I/R lymph drainage,the mucosa thickness and virus height were decreased,but the decrements were significantly lower in TLR4-/-mice;there were no statistically significant differences in the levels of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),endotoxin,and HMGB1 between the TLR4-/-and the WT mice injected with normal lymph or endotoxin.In the mice injected with I/R lymph drainage,the levels of inflammatory factors in the TLR4-/-mice were significantly lower than those in the WT mice [TNF-α:(28.637 ±5.166) pg/ml vs.(41.917 ±8.175) pg/ml,P=0.000;IL-6:(60.900 ±24.729) pg/ml vs.(110.265 ±28.545) pg/ml,P =0.000].In the mice injected with HMGB1,the levels of inflammatory factors in the TLR4-/-mice were significantly decreased compared with those in the WT mice [TNF-α:(20.865 ± 6.464) pg/ml vs.(31.059 ± 6.204) pg/ml,P=0.004;IL-6:(36.268 ±8.977) pg/ml vs.(76.677 ± 14.099) pg/ml,P=0.000].Conclusions The concentrations of endotoxin and HMGB1 are significantly increased during intestinal I/R in rats.After injection of I/R lymph drainage,endotoxin,and HMGB1,the levels of inflammatory factors and HMGB1 in the mice injected with I/R lymph drainage are significantly higher than those in the mice injected with normal lymph;the levels of inflammatory factors and local damage of intestinal mucosa are significantly reduced in the TLR4-/-mice than in the WT mice.The gut-lymph pathway may play a key role in the intestinal I/R injury.

Serum parathyroid hormone as a potential novel biomarker of coronary heart disease.

OBJECTIVE: This study aimed to evaluate the relationships between serum parathyroid hormone (PTH) and coronary heart disease (CHD). METHODS: From July 2011 to February 2013, a total of 79 CHD patients and 94 normal control patients with ages ranging from 25 to 79 years were included in this study. Serum PTH level and common risk factors of CHD (age, gender, cholesterol, glycosylated hemoglobin [HbA1c], blood pressure [BP], history of diabetes, smoking, and body mass index) were investigated. Pearson's correlation and multiple regression analyses were used to evaluate the relationships between serum PTH level and CHD risk factors. All statistical analyses were performed using the SPSS 18.0 software. RESULTS: RESULTS from Pearson's correlation analysis indicated that age, systolic blood pressure (SBP), diastolic blood pressure (DBP), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-c), HbA1c, history of smoking, and serum PTH level were risk factors for CHD (all p<0.05). Serum PTH levels were positively correlated with DBP (r=0.256, p=0.010) and HbA1c (r=0.223, p=0.003), while not being related to other risk factors of CHD (all p>0.05). Multiple linear regression analysis showed that SBP, DBP, LDL-c, and HDL-c may be important determinants of CHD (all p<0.05). Further, serum PTH level is also an independent risk factor for CHD (p<0.001). CONCLUSION: Our results provide evidence that serum PTH level may be involved in the pathogenesis of CHD. Thus, PTH could be used as an important biomarker in the diagnosis of CHD.