Expression of FMD virus-like particles in yeast Hansenula polymorpha and immunogenicity of combine with CpG and aluminum adjuvant

Background@#Inactivated vaccines are limited in preventing foot-and-mouth disease (FMD) due to safety problems. Recombinant virus-like particles (VLPs) are an excellent candidate for a novel vaccine for preventing FMD, given that VLPs have similar immunogenicity as natural viruses and are replication- and infection-incompetent. @*Objectives@#The 3C protease and P1 polyprotein of type O FMD virus (FDMV) was expressed in yeast Hansenula polymorpha to generate self-resembling VLPs, and the potential of recombinant VLPs as an FMD vaccine was evaluated. @*Methods@#BALB/c mice were immunized with recombinant purified VLPs using CpG oligodeoxynucleotide and aluminum hydroxide gel as an adjuvant. Cytokines and lymphocytes from serum and spleen were analyzed by enzyme-linked immunosorbent assay, enzyme-linked immunospot assay, and flow cytometry. @*Results@#The VLPs of FMD were purified successfully from yeast protein with a diameter of approximately 25 nm. The immunization of mice showed that animals produced high levels of FMDV antibodies and a higher level of antibodies for a longer time. In addition, higher levels of interferon-γ and CD4 + T cells were observed in mice immunized with VLPs. @*Conclusions@#The expression of VLPs of FMD in H. polymorpha provides a novel strategy for the generation of the FMDV vaccine.

Taxonomy of Epimedium (Berberidaceae) with special reference to Chinese species / 中草药·英文版

Epimedii Herba is a commonly used traditional Chinese herbal medicine. Five Epimedium species are included in Chinese Pharmacopoeia and most species of Epimedium are used as Epimedii Herba in practical application. However, as the largest herbaceous genus of the Berberidaceae, Epimedium has many taxonomic controversies which hinder the effective use of Epimedii Herba. This paper reviewed the taxonomic research related to Epimedium, including taxonomic history, taxonomic values of morphological characters, species and distribution, infra-genera taxonomic system and the taxonomic research of Chinese Epimedium. For instance, we recognized Epimedium wushanense and clarified that the species, as described in Flora Reipublicae Popularis Sinicae and Flora of China, actually includes four Epimedium species similar in leaflet shape. In general, it was recognized here that Epimedium comprises 62 species, of which 52 species are distributed in China. For Chinese Epimedium species with the most taxonomic problems, the taxonomic research on the taxa was reviewed and the newest species key was proposed along with proposals for those taxonomic problems needing further resolution. This review is of great implication for the identification, exploration and utilization of Epimedii Herba.

Expression of MAD2L1 in Lung Adenocarcinoma and Its Effect on Immune Microenvironment / 肿瘤防治研究

Objective To investigate the expression of MAD2L1 in lung adenocarcinoma and its effect on the prognosis and immune microenvironment of patients. Methods The difference of MAD2L1 expression in lung adenocarcinoma tissue and normal lung tissue was analyzed by TCGA and GEO database. Survival analysis was carried out to evaluate the prognostic significance of MAD2L1 gene expression in lung adenocarcinoma patients. StarBase database was used to construct miRNA-MAD2L1 regulatory network of lung adenocarcinoma. The relation between the expression of MAD2L1 and immune cell infiltration in lung adenocarcinoma was analyzed by TIMER database. Results The expression of MAD2L1 was up-regulated in lung adenocarcinoma, and the high expression of MAD2L1 was significantly correlated with pathological stage and lymph node metastasis of lung adenocarcinoma. The patients with high expression of MAD2L1 had a poor prognosis. miR-101-3p/MAD2L1 axis was identified as the most potential upstream regulation pathway of MAD2L1 in lung adenocarcinoma. The expression level of MAD2L1 was significantly correlated with tumor immune cell infiltration and immune checkpoint expression. Conclusions MAD2L1 is highly expressed in lung adenocarcinoma, which is related to poor prognosis and tumor immune infiltration. MAD2L1 can be used as a potential target for the treatment of lung adenocarcinoma.

Inhibition of tanshinone ⅡA on lung cancer cell proliferation / 预防医学

Objective@#To investigate the inhibition of tanshinoneⅡA (TanⅡA) on the proliferation of lung cancer cells.@*Methods@#Human lung cancer cell lines A549, SK-MES-1, H446 and H460 were cultured in vitro and treated with TanⅡA at concentrations of 0.3, 0.6, 1.3, 2.5, 5.0, 10.0 mg/mL, while untreated cells served as controls. Cell proliferation was measured by using CCK-8 assay, and apoptosis was measured using flow cytometry, while the expression of apoptosis-related proteins was determined using Western blotting. The apoptotic rate of lung cancer cells was compared between Tan ⅡA-treated cells and untreated cells.@*Results@# Tan ⅡA inhibited lung cancer cell proliferation in a time-dependent and concentration-dependent manner, and the survival rates of A549, SK-MS-1, H446 and H460 cells reduced with the concentration of TanⅡA (t=4.503, 2.114, 2.103 and 3.567; all P<0.05) and the duration of TanⅡA treatment (t=5.189, 3.079, 3.023 and 3.845; all P<0.05). The 48 h half maximal inhibitory concentrations (IC50 values) of TanⅡA were (1.18±0.12), (0.78±0.08), (1.55±0.16) and (1.27±0.14) mg/mL against A549, SK-MES-1, H446 and H460 cells, respectively. Following 48 h treatment with Tan ⅡA at a concentration of 2.5 mg/mL, the apoptotic rates of A549, SK-MS-1, H446 and H460 cells were (34.97±3.78)%, (37.62±2.48)%, (18.27±2.98)% and (19.17±2.30)%, which were significantly significantly higher than those of untreated cells [(4.86±0.36) %, (3.21±0.48) %, (3.25±0.26)% and (2.66±0.19)%, all P<0.05]. Reduced Akt1 protein expression, elevated Bax/Bcl-2 expression ratio, and elevated caspase-9 and caspase-3 protein expression were detected in lung cancer cells treated with 2.5 mg/mL TanⅡA for 48 h relative to untreated cells@*Conclusion@#TanⅡA may inhibit lung cancer cell proliferation in a time- and concentration-dependent manner via the Bax/Bcl-2/caspase-9/caspase-3 pathway.

3D printed guide template used in osteotomy for malunion of tibial fracture / 中华创伤骨科杂志

Objective:To evaluate the personalized 3D printed guide template used in the osteotomy for malunion of tibial fracture.Methods:A retrospective analysis was conducted of the 30 patients who had been treated for malunion of tibial fracture at Department of Orthopaedics, The First Affiliated Hospital to Zhengzhou University from January 2010 to January 2018. Of them, 15 used a personalized 3D printed guide template in the osteotomy (3D printing group). They were 9 males and 6 females, with an age of 46.3 year±8.2 years. The fracture malunion was located in the upper and middle tibia in 11 cases, in the lower tibia in 4 cases, on the left side in 6 cases and on the right side in 9 ones. There were 8 cases of varus deformity and 7 ones of valgus deformity. Their preoperative fracture deformity angle was 24.3°±5.5°. The other 15 patients were treated with conventional surgery (conventional group). They were 10 males and 5 females, with an age of 47.1 years±6.0 years. The fracture was located in the upper and middle tibia in 12 cases, in the lower tibia in 3 cases, on the left side in 5 cases and on right side in 10 cases. There were 7 cases of varus deformity and 8 ones of valgus deformity. Their preoperative fracture deformity angle was 22.5°±5.4°. The 2 groups were compared in terms of preoperative baseline data, operation time, intraoperative blood loss and postoperative recovery of the alignment of lower limb.Results:There were no significant differences in the preoperative baseline data between the 2 groups, showing comparability ( P>0.05). The 3D printing group was followed up for an average of 12 months while the conventional group for an average of 10 months. The operation time for the 3D printing group was significantly shorter than that for the conventional group(102.2 min±13.0 min versus 137.9 min ±10.5 min), the intraoperative blood loss for the former significantly less than that for the latter (77.3 mL ± 39.7 mL versus 163.3 mL ± 35.2 mL), and the postoperative malunion angle in the former significantly smaller than that in the latter (1.9°±0.4° versus 3.2°±0.9°) (all P< 0.05). The last follow-ups revealed no implant failure or re-malunion but fine healing of the osteotomy sites and good recovery of the alignment of lower limb in the 2 groups. Conclusion:A personalized 3D printed guide template used in the osteotomy for malunion of tibial fracture is an effective aid because it can facilitate precise osteotomy, reduce operation time and intraoperative blood loss and help correct the alignment of lower limb, leading to good short-term surgical outcomes.

Analysis of changes in coronary arterial microcirculation index and its influencing factors in elderly patients with hypertension / 中华老年医学杂志

Objective@#To analyze the changes in coronary arterial microcirculation index and its possible influencing factors in elderly patients with hypertension.@*Methods@#In the retrospective case-control study, 36 elderly patients with hypertension in our hospital from May 2016 to April 2017 and meeting inclusion criteria were recruited as the hypertension group, and 36 matched non-hypertensive elderly patients during the same time were considered as the non-hypertensive group.Patients in the two groups underwent coronary angiography and percutaneous coronary function tests for the coronary heart disease symptoms such as chest pain and chest tightness.The changes of coronary microcirculation index were observed, and its possible influencing factors were analyzed in combination with other test results.@*Results@#The low-density lipoprotein level was lower, and plasma viscosity, erythrocyte sedimentation rate, serum high-sensitivity C-reactive protein and 24-h urinary protein were higher in the hypertension group than in the non-hypertensive group(P<0.05). The parameters were higher in the hypertension group versus in control group as follows: Left ventricular ejection fractions(LVEF), [(74.82±6.71 )% vs.(71.56±4.93 )%], Left ventricular mass[(189.72±33.40)g vs.(174.38±29.11)g], Left ventricular mass index[(101.70±16.25)g/cm2 vs.(89.68±10.46)g/cm2](t=2.077, 3.731 and 4.676, P<0.05 or P<0.01), and the proportion of patients with decreased LVEF(52.8%, 19 cases vs.27.8% 10 cases, χ2=4.676). Multivariate logistic regression analysis showed that low-density lipoprotein(OR=2.887, 95%CI: 1.754~8.193, P=0.035), plasma viscosity(OR=4.075, 95%CI: 1.629~7.386, P=0.001), erythrocyte sedimentation rate(OR=3.534, 95%CI: 1.293~6.730, P=0.001), serum high-sensitivity C-reactive protein(OR=2.821, 95%CI: 1.546~7.386, P=0.001), 24-h urinary protein(OR=3.712, 95%CI: 1.683~7.954, P=0.026), left ventricular mass index(OR=3.254, 95%CI: 1.286~6.735, P=0.004)and reduced left ventricular diastolic function(OR=5.069, 95%CI: 1.386~7.254, P=0.007)were the risk factors for abnormal changes in coronary microcirculation resistance index.@*Conclusions@#Elderly hypertensive patients have abnormal coronary microcirculation resistance index, and the intervention should be strengthened from the aspects of blood lipid, plasma viscosity, , inflammation, renal function and cardiac function, so as to improve coronary function and reduce adverse cardiovascular events.

Analysis of changes in coronary arterial microcirculation index and its influencing factors in elderly patients with hypertension / 中华老年医学杂志

Objective To analyze the changes in coronary arterial microcirculation index and its possible influencing factors in elderly patients with hypertension.Methods In the retrospective case-control study,36 elderly patients with hypertension in our hospital from May 2016 to April 2017 and meeting inclusion criteria were recruited as the hypertension group,and 36 matched nonhypertensive elderly patients during the same time were considered as the non-hypertensive group.Patients in the two groups underwent coronary angiography and percutaneous coronary function tests for the coronary heart disease symptoms such as chest pain and chest tightness.The changes of coronary microcirculation index were observed,and its possible influencing factors were analyzed in combination with other test results.Results The low-density lipoprotein level was lower,and plasma viscosity,erythrocyte sedimentation rate,serum high-sensitivity C-reactive protein and 24-h urinary protein were higher in the hypertension group than in the non-hypertensive group(P＜0.05).The parameters were higher in the hypertension group versus in control group as follows:Left ventricular ejection fractions (LVEF),[(74.82 ±-6.71) ％ vs.(71.56 ±-4.93) ％],Left ventricular mass[(189.72 ± 33.40) g vs.(174.38 ± 29.11) g],Left ventricular mass index [(101.70 ± 16.25)g/cm2 vs.(89.68 ± 10.46) g/cm2] (t =2.077,3.731 and 4.676,P ＜0.05 or P ＜0.01),and the proportion of patients with decreased LVEF (52.8％,19 cases vs.27.8％ 10 cases,x2 =4.676).Multivariate logistic regression analysis showed that low-density lipoprotein(OR =2.887,95 ％ CI:1.754～ 8.193,P =0.035),plasma viscosity(OR =4.075,95％ CI:1.629 ～ 7.386,P =0.001),erythrocyte sedimentation rate (OR =3.534,95％ CI:1.293 ～ 6.730,P =0.001),serum highsensitivity C-reactive protein (OR =2.821,95 ％ CI:1.546 ～ 7.386,P =0.001),24-h urinary protein (OR=3.712,95％CI:1.683～7.954,P =0.026),left ventricular mass index(OR =3.254,95％CI:1.286 ～ 6.735,P =0.004)and reduced left ventricular diastolic function(OR =5.069,95 ％ CI:1.386 ～7.254,P=0.007)were the risk factors for abnormal changes in coronary microcirculation resistance index.Conclusions Elderly hypertensive patients have abnormal coronary microcirculation resistance index,and the intervention should be strengthened from the aspects of blood lipid,plasma viscosity,,inflammation,renal function and cardiac function,so as to improve coronary function and reduce adverse cardiovascular events.

Co-occurrence of t(8;21)(q22;q22) and t(9;22)(q34;q11) in a case with chronic myelogenous leukemia / 中华医学遗传学杂志

OBJECTIVE@#To delineate laboratory and clinical characteristics of a case with chronic myelogenous leukemia (CML) and co-occurrence of t(9;22)(q34;q11) and t(8;21)(q22;q22).@*METHODS@#The patient was subjected to cytogenetic, molecular, morphological and immunophenotypic analyses.@*RESULTS@#Cytogenetic analysis revealed presence of t(8;21)(q22;q22) in addition to t(9;22)(q34;q11) in the patient. Chimeric BCR/ABL and AML1/ETO genes were detected by fluorescence in situ hybridization (FISH). Transcripts of BCR/ABL210 and AML1/ETO fusion genes were detected by relative quantity PCR. Morphological study suggested that the patient was at the chronic phase of CML. No significant immunophenotypic abnormality was detected by flow cytometry.@*CONCLUSION@#Co-occurrence of t(8;21)(q22;q22) and t(9;22)(q34;q11) is rare in CML. Only 5 similar cases have been described previously. This case suggested that chromosomal alterations may precede morphological, flow cytometric and clinical changes and accelerate progression of the disease.

Clinical characteristics of high-grade B-cell lymphomas with rearrangement of MYC, bcl-6 and bcl-2 / 中华病理学杂志

Objective@#To investigate the clinicopathologic features of patients with high-grade B-cell lymphomas (HGBL) that have rearrangements of MYC, bcl-6 and bcl-2.@*Methods@#One hundred and fifty-eight B-cell lymphomas patients from Institute of Hematology and Blood Diseases Hospital from January 2016 to April 2017 were detected by fluorescence in situ hybridization (FISH) with double color split-apart probes.@*Results@#Among 158 B-cell lymphomas, 3 cases with MYC, bcl-2 and bcl-6 rearrangements were identified, 1 of which also had CCND1/IgH translocation. All three patients were of older age, with poor prognostic parameters, multiple organs involvements, elevated LDH and advanced-tumor stage. Two of the three patients were treated with high-intensity chemotherapy and had no remission with an overall survival of 9 months and 11 months respectively. One patient had follow-up with no treatment. Histologically, all three cases showed a spectrum of morphologic features. Although initially categorized as lymphoblastic lymphoma, diffuse large lymphoma and mantle cell lymphoma respectively, two cases were associated with germinal center B-cell (GCB) immunophenotype and 1 case with non-GCB immunophenotype. They had a high proliferation index as assessed by immunostaining for Ki-67 (60%-90%).@*Conclusions@#MYC+ bcl-2+ bcl-6+ HGBL is an aggressive disease with multiple organ involvement, high serum LDH levels, advanced stage disease, poor prognosis and shorter patient survival. The diagnosis should be made by histopathology combined with FISH analysis. Its separation from other types of B cell large cell lymphoma is of clinical importance.

The Efficacy of Etanercept Combined Leflunomide to Treat Ankylosing Spondylitis and Its Safety / 现代生物医学进展

Objective:To explore the efficacy of etanercept combined leflunomide to treat ankylosing spondylitis (AS) and its safety.Methods:90 cases with AS patients admitted in our hospital from June 2016 to January 2015 were selected.The patients were divided into observation group and control group by random number table method,45 cases in each group.Observation group was treated with etanercept combined with leflunomide,the control group were treated by leflunomide only.Morning stiffness time,AS activity index (BASDAI),AS measurement index (BASMI) were recorded before and after treatment in two groups,the total effective rate and adverse reaction were compared between the two groups.Results:After treatment,the BASDAI and morning stiffness time,BASMI of two groups of patients compared with before treatment were decreased,and the indexes above of the observation group were significantly lower than the control group,the difference was statistically significant (P＜0.05);The effective rate of the observation group was significantly higher than that of the control group,the difference was statistically significant (P＜0.05);During the course of treatment,the main adverse reactions occurred in patients with liver function damage,diarrhea and allergic skin rash,but there was no significant difference in the incidence of adverse reactions between the two groups (P＞0.05).Conclusion:Etanercept combined with leflunomide can obtain ideal effect,and low incidence rate of adverse reaction,it is worth popularizing in clinical use.

Advances in percutaneous pulmonary valve implantation / 中华胸心血管外科杂志

Percutaneous pulmonary valve implantation (PPVI) is an exciting growing field in cardiovascular medicine.With the sophisticated improvement of biomedical engineering,development of PPVI offers people a new way of less invasive techniques to treat these groups of patients with pulmonary valvular regurgitation over the past several years.New stent designs and operative and interventional hybrid approaches are under investigation.This nonsurgical approach has been proven to be feasible and holdpromise,although many obstacles still exist.At the present time,PPVI has been widely used in clinical in the United States and European countries,but in our country,only a small number of cases reports have been published.With wider application and development of PPVI,cumulative experiences and long-term follow-up of PPVI therapies,better criteria will be established for patient election.Complications will be reduced and safer and more effective treatment results will be achieved.PPVI therapies will be applied to a larger subset of patients and become an important alternative to future conventional pulmonary valve replacement.

Analysis of risk factors for postoperative atrial fibrillation in elderly patients with coronary heart disease after off-pump coronary artery bypass grafting surgery / 中华老年医学杂志

Objective To investigate certain risk factors for postoperative atrial fibrillation (AF) in elderly patients with coronary heart disease after off-pump coronary artery bypass grafting (OPCAB)surgery in order to provide a basis for the prevention and treatment of AF after OPCAB.Methods A total of 139 elderly patients with coronary heart disease who had undergone OPCAB surgery in our hospital were collected as research subjects and divided into the AF group and the nonAF group according to the occurrence of AF after operation.The patients' general information and clinical data were retrospectively analyzed.Risk factors for AF after OPCAB surgery in elderly patients with coronary heart disease were investigated.Results The incidence of AF after OPCAB was 15.8％ in elderly patients with coronary heart disease.There were no statistical differences in gender,history of diabetes,history of myocardial infarction,preoperative β-blocker usage,number of coronary artery lesions,or operation time between the AF group and the non-AF group (P＞0.05).The percentage of patients who were of older age (t=9.960) or had a left ventricular ejection fraction ≥40％ (x2=4.942),a left atrial diameter ≥40 mm (x2 =4.491),a history of hypertension (x2 =12.357),dopamine medication after operation (x2 =8.511),or a bypass vessel count≥ 3 (x2 =5.385) was higher in the AF group than in the non-AF group (all P＜0.05).Logistic regression analysis showed that age,history of hypertension,dopamine medication after operation,left atrial diameter and bypass vessel count were the risk factors for AF after OPCAB surgery in elderly patients with coronary heart disease (OR 3.080,2.435,2.465,3.593,and 1.921,respectively,P＜0.05 for all).Conclusions The incidence of AF after OPCAB is high in elderly patients with coronary heart disease and is affected by many risk factors.These risk factors should be assessed before surgery so that appropriate prevention measures can be taken.

Clinical Study onDu-Huo Ji-ShengDecoction with Low Frequency Ultrasonics in Treatment of Knee Osteoarthritis / 世界科学技术-中医药现代化

This study was aimed to observe the clinical effect of Du-Huo Ji-Sheng (DHJS) decoction with low frequency ultrasonics in treatment of knee osteoarthritis (KOA) by evaluating the visual analogue scale (VAS), Lysholm score and the expression levels of transforming growth factor-β (TGF-β) and matrix metalloproteinase-3 (MMP-3). A total of 60 KOA outpatients were randomly divided into the treatment group and the control group, with 30 cases in each group. Patients in the treatment group were treated with DHJS decoction and low frequency ultrasonics therapy, while patients in the control group were treated with ibuprofen and low frequency ultrasonics. The treatment was given once a day for two weeks. The pretreatment and posttreatment VAS, Lysholm score and expression levels of TGF-β and MMP-3, which were detected by ELISA, were recorded. The results showed that compared with the control group, DHJS decoction with low frequency ultrasonics therapy could alleviate pain as well as improve the activity of joint function among KOA patients. It could also increase the content of TGF-βin serum, and suppress the level of MMP-3. It was concluded that low frequency ultrasonics could enhance the transdermal absorption of DHJS decoction, relieve the clinical symptoms of KOA patients.

Phylogenetic and Bioinformatics Analysis of Replicase Gene Sequence of Cucumber Green Mottle Mosaic Virus / 病毒学报

The replicase genes of five isolates of Cucumber green mottle mosaic virus from Jiangsu, Zhejiang, Hunan and Beijing were amplificated, sequenced and analyzed. The similarities of nucleotide acid sequences indicated that 129 kD and 57 kD replicase genes of CGMMV-No. 1, CGMMV-No. 2, CGMMV-No. 3, CGMMV-No. 4 and CGMMV-No. 5 were 99.64% and 99.74%, respectively. The similarities of 129 kD and 57 kD replicase genes of CGMMV-No. 1, CGMMV-No. 3 and CGMMV-No. 4 were 99.95% and 99.94%, while they were lower between CGMMV-No. 2 and the rest of four reference sequences, just from 99.16% to 99.27% and from 99.04% to 99.18%. All reference sequences could be divided into six groups in neighbor-joining (NJ) phylogenetic trees based on the replicase gene sequences of 129 kD, 57 kD protein respectively. CGMMV-No. 1, CGMMV-No. 3 and CGMMV-No. 4 were clustered together with Shandong isolate (Accession No. KJ754195) in two NJ trees; CGMMV-No. 5 was clustered together with Liaoning isolate (Accession No. EF611826) in two NJ trees; CGMMV-No. 2 was clustered together with Korea watermelon isolate (Accession No. AF417242) in phylogenetic tree of 129 kD replicase gene of CGMMV; Interestingly, CGMMV-No. 2 was classified as a independent group in phylogenetic tree of 57 kD replicase gene of CGMMV. There were no significant hydrophobic and highly coiled coil regions on 129 kD and 57 kD proteins of tested CGMMV isolates. Except 129 kD protein of CGMMV-No. 4, the rest were unstable protein. The number of transmembrane helical segments (TMHs) of 129 kD protein of CGMMV-No. 1, CGMMV-No. 2, CGMMV-No. 3 and CGMMV-No. 5 were 6, 6, 2 and 4, respectively, which were 13, 13 and 5 on the 57 kD protein of CGMMV-No. 2, CGMMV-No. 4 and CGMMV-No. 5. The glycosylation site of 129 kD protein of tested CGMMV isolates were 2, 4, 4, 4 and 4, and that of 57 kD protein were 2, 5, 2, 5 and 2. There were difference between the disorders, globulins, phosphorylation sites and B cell antigen epitopes of 129 kD and 57 kD proteins of tested CGMMV isolates. The current results that there was no significant difference between the replicase gene sequences, it was stable and conservative for intra-species and clearly difference for inter-species. CGMMV-No. 1, CGMMV-No. 3, CGMMV-No. 4 and CGMMV-No. 5 had. a close genetic relationship with Shandong and Liangning isolates (Accession No. KJ754195 and EF611826), they are potentially originate from the same source. CGMMV-No. 2 was closer with Korea isolate. High sequence similarity of tested samples were gathered for a class in phylogenetic tree. It didn't show regularity of the bioinformatics analysis results of 129 kD and 57 kD proteins of tested CGMMV isolates. There was no corresponding relationship among the molecular phylogeny and the bioinformatics analysis of the tested CGMMV isolates.

Combined treatment with erythropoietin and granulocyte colony-stimulating factor enhances neovascularization and improves cardiac function after myocardial infarction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Erythropoietin (EPO) and granulocyte colony-stimulating factor (G-CSF) are both potential novel therapeutics for use after myocardial infarction (MI). However, their underlying mechanisms remain unclear and the efficacy of monotherapy with EPO or G-CSF is also controversial. Therefore, we investigated the effects of combined treatment with EPO and G-CSF on neovascularization and cardiac function in post-infarction rats and explored the potential mechanisms.</p><p><b>METHODS</b>Four groups of rats were used: control (saline injection after MI, i.h.), EPO (a single dose of 5 000 IU/kg after MI, i.h.), G-CSF (a dose of 50 µg× kg(-1)× d(-1) for 5 days after MI, i.h.), and both EPO and G-CSF (EPO+G-CSF, using the same regiment as above). Cardiac function was assessed by echocardiography before and 1 day, 7 days, 14 days and 21 days after MI. CD34(+)/Flk-1(+) cells in the peripheral blood were evaluated by flow cytometry before and 3 days, 5 days and 7 days after MI. The infarct area and angiogenesis in the peri-infarct area were analyzed. The mRNA and protein expression of vascular endothelial growth factor (VEGF) and stromal-derived factor-1a (SDF-1α) in the peri-infarct area were detected by real-time quantitative RT-PCR and Western blotting.</p><p><b>RESULTS</b>Compared with the control and monotherapy groups, the EPO+G-CSF group had significantly increased CD34(+)/Flk-1(+) endothelial progenitor cells (EPCs) in the peripheral blood (P < 0.05), up-regulated VEGF and SDF-1α levels in the peri-infarct region (P < 0.05), enhanced capillary density (P < 0.05), reduced infarct size (P < 0.05) and improved cardiac structure and function (P < 0.05). G-CSF alone did not dramatically increase EPCs in the peripheral blood, enhance capillary density in the peri-infarct area or reduce infarct size compared with the control group.</p><p><b>CONCLUSIONS</b>Combined treatment with EPO and G-CSF increased EPCs mobilization, up-regulated VEGF and SDF-1α levels in the post-infarction microenvironment, subsequently enhanced neovascularization in the peri-infarct region and reduced infarct size. All factors contributed to its beneficial effects on cardiac function in post-infarction rats.</p>

Construction of a capsular tissue-engineered ureteral stent seeded with autologous urothelial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the feasibility of constructing a capsular poly L-lactic acid (PLLA) ureteral stent seeded with autologous urothelial cells using tissue engineering methods.</p><p><b>METHODS</b>The capsular ureteral stent was constructed by subcutaneously embedding PLLA ureteral stent in the back of beagles for 3 weeks to induce the formation of connective tissue on the surfaces. After decellularization of the stent, the expanded autologous urothelial cells were seeded on the stent. The surface structure and cell adhesion of the stent were observed using HE staining, scanning electron microscope (SEM) and immunocytochemical staining. MTT assay was used to evaluate urothelial cell proliferation on the capsular PLLA ureteral stent and on circumferential small intestinal submucosa graft.</p><p><b>RESULTS</b>HE staining and VIII factor immunohistochemistry revealed numerous capillaries in the connective tissue encapsulating the stent without obvious local inflammatory response. The results of SEM and immunocytochemical staining showed that the capsule contained rich collagenic fibers forming three-dimensional structures, and the seeded autologous urothelial cells could adhere and well aligned on the surface. MTT assay showed normal growth of the cells on the stent as compared with the cells grown on circumferential small intestinal submucosa graft.</p><p><b>CONCLUSION</b>The capsular PLLA ureteral stent allows adhesion and proliferation of autologous urothelial cells and shows a potential in applications of constructing tissue-engineered ureter.</p>

Correlation study between femoral distal medial torsion and patellofemoral joint malalignment / 中国医师进修杂志

ObjectiveTo investigate the correlation between femoral distal medial torsion and patellofemoral joint malalignment and analyze the causes of patellofemoral joint disorders,which provide the new theory with clinical treatment.MethodsFrom May 2007 to June 2009,124 knees(95 cases) with patellofemoral joint disorders were enrolled in this study randomly.Each knee was scanned with CT in dynamic 20° -30° knee flax position.Femoral distal medial torsion angle (FMTA),patellar congruence angle (CA) and patellar tilt angle(PTA) were measured.The correlation between FMTA and CA or PTA was analyzed.Results FMTA ＜ 5° in 25 knees,≥5° in 99 knees,6 knees with trochlear dysplasia who were excluded.FMTA in 93 knees was 16.06° ± 5.68°,CA was 16.40° ± 5.48° and PTA was 19.59° ± 3.32°.The positive correlation was found between FMTA and CA when FMTA ＞ 10°through scatter diagram analysis (r =0.709,P ＜ 0.05 ).The positive correlation was found between FMTA and PTA when FMTA ＞10°( r =0.652,P ＜ 0.05),the positive trend declined when FMTA ＞ 27°.ConclusionsFemoral distal medial torsion is an important risk factor of patellofemoral joint malalignment.When FMTA ＞ 10°,FMTA and CA,PTA has positive correlation,but the positive tend between FMTA and PTA declines when FMTA ＞ 27°.

Genetic Analysis of the P1 Region of Human Enterovirus 71 Strains and Expression of the 55 F StrainVP1 Protein / 中国病毒学·英文版

Enterovirus 71 (EV71) is a member of the Entero-virus genus of the Picornaviridae family and is the major cause of Hand,foot,and mouth disease (HFMD) in children.Different strains from Gansu were cloned and the P1 protein was sequenced and analysed.Results indicate that there are three kinds of EV71 infections prevalent in Gansu.The VP1 protein from one of these strains,55F,was expressed.The recombinant protein was expressed with high level and reacted specifically with the EV71 patient antibody,the recombinant protein was also applied to raise antiserum in rabbits and after the fourth injection a high titer of antiserum was detected by ELISA assay.These data are useful for further clarification of prevalent EV71 strains in the north of China at the molecular level and provide a basis for EV71 diagnosis.

Salvation for failure of lateral retinacular release / 中国医师进修杂志

Objective To investigate a simple effective way to salvage failure of lateral retinacular release. Methods From March 2007 to May 2009,there were 23 knees of 23 cases for failure after lateral retinacular. All the patients had patellofemoral joint malatignment in some extend,whose patellofemoral joint CT scanned in dynamic. Before curing,the index of patellar tilting angle (PTA) was 19.6° ±5.7° , congruence angle(CA) was 23.6° ± 12.5° , patellar femoral index(PFI)was 2.7 ± 0.8, and hospital special score (HSS) knee score was (74.0 ± 9.2) points,then put on elastic knee sleeves for arranging the malalignment of patellofemoral joints, coincidentally, rehabilitation of vastus medians administered. Results All the patients were followed up for 6 - 26 (11.0 ± 2.3 ) months, 22 cases of 24 knees gained the excellent or good results,1 case had to experience surgery again. After curing, PTA was 11.2° ±3.7°,CA was 6.4° ±7.8°, and PFI was 1.1 ±0.3,escalated HSS knee score was (87.0 ±6.5) points, in these excellent or good patellofemoral joints. Significant difference showed apparently in the cohort through curing (P <0.05). Conclusion Administering elastic knee sleeves along with correct rehabilitation is a simple effective no-incisive cure for salvaging failure of lateral retinacular release.

Prokaryotic Expression and Potential Application of the Truncated PCV-2 Capsid Protein / 中国病毒学·英文版

Three pairs of specific primers were designed to amplify the F2-1, F2-2 and XF2-2 truncated sequences of ORF2 which encodes the capsid protein of porcine circovirus type 2 (PCV-2). The F2-1 sequence had most of the NLS region of ORF2, but the F2-2 and XF2-2 genes had the NLS region deleted. Truncated genes were subcloned into pET-32a(+) vectors to construct recombinant fusion expression vectors. The vectors were then transformed into Rosetta(DE3) E. coli and expressed by induction of IPTG. Expressed proteins were detected by western blotting and ELISA. The protein with best immunoreactivity was confirmed and selected, then utilized to inoculate SPF rabbits to prepare polyclonal antibodies. The protein and prepared polyclonal antibody were utilized to detect sera samples against PCV-2 from Shandong province and PCV-2 particles in PK-15 cells. In our study, three recombinant fusion proteins were successfully obtained, and the molecular weights of fusion proteins were 35.9 kDa, 33.6 kDa and 38.6 kDa respectively detected by SDS-PAGE. All of the proteins showed positive reaction with anti-PCV-2 antisera, and His-XF2-2 showed better immunoreactivity than the others. The protein of His-XF2-2 was coated as antigen in ELISA to detect the seroprevalence of PCV-2 in certain districts of Shandong province, the seropositivity rate was 27.7 % (73/264). Specific fluorescence and positive signals for PCV-2 could be detected in PK-15 cells inoculated with PCV-2 with the participation of prepared antibodies against His-XF2-2 in IFA and IPMA. Experimental results indicated that the truncated PCV-2 ORF2 gene containing most of the NLS region was successfully expressed in E. coli, and His-XF2-2 was demonstrated to have better immunoreactivity with anti-PCV-2 antisera than the other two fusion proteins. His-XF2-2 and prepared polyclonal antibodies against it had a satisfactory capability in detecting PCV-2 infection.