Application Value of Sevoflurane Combined with Brachial Plexus Block in Children with Humeral Fracture Surgery and its Effect on Hemodynamics: A Clinical and Anatomical Study

SUMMARY: To observe the effect of sevoflurane combined with brachial plexus block (BPB) in children with humeral fracture surgery and its effect on hemodynamics. 84 children who received surgical treatment of humeral fracture in our hospital from September 2019 to September 2022 were selected. According to different anesthesia methods, the children were divided into control group and study group. The control group only received laryngeal mask sevoflurane; the study group received laryngeal mask sevoflurane combined with BPB. The operation situation, hemodynamic indexes, stress level, pain and adverse reactions of children was observed. The postoperative awakening time in the study group was lower than control group, the postoperative pain onset time in the study group was higher than control group (P0.05). Postoperative 2h, the levels of serum cortisol, b-endorpin, norepinephrine and epinephrine in the study group were lower than control group (P0.05). Sevoflurane combined with BPB is helpful to shorten the postoperative awakening time of children with humeral fracture, reduce the degree of postoperative pain, improve hemodynamics, and reduce stress response, and has good safety.

El objetivo fue observar el efecto del sevoflurano combinado con bloqueo del plexo braquial (BPB) en niños con cirugía de fractura de húmero y su efecto sobre la hemodinámica. Se seleccionaron 84 niños que recibieron tratamiento quirúrgico de fractura de húmero en nuestro hospital desde septiembre de 2019 hasta septiembre de 2022. Según diferentes métodos de anestesia, los niños se dividieron en grupo control y grupo de estudio. El grupo control solo recibió sevoflurano en mascarilla laríngea; el grupo de estudio recibió sevoflurano con mascarilla laríngea combinado con BPB. Se observó la situación operatoria, índices hemodinámicos, nivel de estrés, dolor y reacciones adversas de los niños. El tiempo hasta el despertar postoperatorio en el grupo de estudio fue menor que el del grupo control, el tiempo de aparición del dolor postoperatorio en el grupo de estudio fue mayor que el del grupo control (P0,05). A las 2 horas postoperatorias, los niveles séricos de cortisol, β-endorfina, norepinefrina y epinefrina en el grupo de estudio fueron más bajos que los del grupo control (P 0,05). El sevoflurano combinado con BPB es útil para acortar el tiempo de despertar del posoperatorio de los niños con fractura de húmero, reduce el grado de dolor postoperatorio, mejora la hemodinámica y reduce la respuesta al estrés, además de tener buena seguridad.

Correlation between tumor necrosis factor-α and interleukin-1β in exhaled breath condensate and pulmonary function in chronic obstructive pulmonary disease / 中国热带医学

@#Abstract: Objective To explore the correlation between lung function in patients with chronic obstructive pulmonary disease (COPD) and tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) levels in exhaled breath condensate (EBC), and to provide a convenient methodological basis for the diagnosis and treatment of COPD and the determination of its efficacy. Methods A total of 81 COPD patients and 40 healthy controls were selected from the respiratory department of the Fourth Affiliated Hospital of Guangzhou Medical University from August 2020 to February 2022 as the research subjects. The COPD patients were divided into 41 cases in the acute exacerbation group and 40 cases in the remission group according to their status. All participants underwent lung function detection, venous blood and EBC collection, and the levels of TNF-α and IL-1β in EBC and venous blood were analyzed by enzyme-linked immunosorbent assay (ELISA), and correlation analysis was performed by Pearson correlation analysis method. Results The levels of TNF-α and IL-1β in EBC of in the acute exacerbation group, the healthy control group, the remission group were (5.16±0.18) pg/μL and (7.75±0.27) pg/μL, (2.66±0.31) pg/μL and (2.41±0.24） pg/μL, (3.61±0.29) pg/μL and (3.17±0.38) pg/μL, respectively. Compared with the healthy control group, the levels of TNF-α and IL-1β in EBC in the COPD acute exacerbation group were significantly higher than those in the healthy control group and the COPD remission group (F=9.451, 8.217, P<0.001). Serum tests were consistent with this result. Correlation analysis showed that the levels of TNF-α and IL-1β in EBC were significantly positively correlated with the level of serum inflammation levels (P<0.001), while significantly negatively correlated with lung function (P<0.001).  Conclusions TNF-α and IL-1β in EBC are potential biomarkers of inflammation in patients with COPD, and their detection can be used to effectively assess lung function in patients with COPD. 

Characteristics and management mechanism of imported malaria from overseas during the malaria elimination phase in Wuhan City / 公共卫生与预防医学

Objective To analyze the characteristics of imported malaria epidemic from overseas in Wuhan, to explore the management mechanism of on-site cases, and to accumulate experience for the treatment of imported malaria in large cities after malaria elimination. Methods The epidemiological data on imported malaria from abroad during the period of malaria elimination (2010-2019) in Wuhan were collected. The gender, age and severe illness-related factors of the cases were analyzed. Based on the characteristics of the epidemic and the current situation of prevention and control, the content and experience of the “Municipal-District 24-7” case mechanism were discussed. Results The medical resources in Wuhan were the best in the central region, resulting in a large number of imported malaria cases, with a total of 474 cases reported from 2010 to 2019 (40.79% of the total number of cases in Hubei Province), including 359 cases of falciparum malaria, 36 severe cases and one death (the death rate was 0.28%). The patients were mainly young and middle-aged men aged 20 to 49 years old (97.26%). There were many referral cases (40.30%), and there was no seasonal clustering of cases reported. The undiagnosed proportion at the first visit was 44.85%, and the time of attack-diagnosis was 4 days or more in 61.00% of cases. The occurrence of severe cases was related to unconfirmed diagnosis at the first visit (χ2=35.46, P<0.001) and attack-diagnosis time (Z=-6.49, P<0.001). Conclusion Imported malaria occurs frequently in Wuhan, mainly falciparum malaria. However, “Municipal-District 24-7” case mechanism has effectively curbed the occurrence of severe and death cases and provided valuable experience for case management in similar cities in China.

Percutaneous thermal ablation combined with simultaneous transarterial chemoembolization for hepatocellular carcinoma ≤5 cm

Background/Aim: Percutaneous thermal ablation combined with transarterial chemoembolization (TACE) becomes a treatment option for unresectable hepatocellular carcinoma (HCC). This study aims to investigate the safety and feasibility of percutaneous thermal ablation combined with simultaneous TACE for patients with HCC ≤ 5 cm. Materials and Methods: From June 2010 to February 2017, a total of 280 patients with HCC ≤ 5 cm who underwent percutaneous thermal ablation combined with simultaneous TACE were included in our study. Their clinical data were collected and analyzed. Results: Major complications occurred in five cases (1.8%). The complete necrosis rate was 91.9%. The median overall survival (OS) was 66.5 months (95% confidence interval [CI] = 57.7–75.2). The OS rates in 1-, 3-, 5-, and 7-year were 96.7%, 76.0%, 59.7%, and 31.1%, respectively. Tumor size (hazard ratio = 1.826; 95% CI = 1.131–2.947; P = 0.014) was considered as independent prognostic factors of long-term survival. Conclusion: Percutaneous thermal ablation combined with simultaneous TACE is a safe and effective treatment for HCC ≤ 5 cm.

Socket shield technique for peri-implant tissue preservation in esthetics zone / 中国组织工程研究

BACKGROUND: Absorption of the alveolar bone subsequent to tooth extraction is a long-term process, especially in the anterior tooth region where the buccal part of the ridge undergoes a prominent remodeling process after tooth extraction. Socket shield technique (SST) recently reported may be a new choice for peri-implant tissue preservation. OBJECTIVE: To give an overview on the principle, interface basis and application of SST in implant dentistry. METHODS: The first author searched the PubMed, Ovid, CNKI, VIP and WanFang databases for relevant articles published from January 1990 to October 2017. The keywords were "socket shield technique;root-membrane technique; root-implant; implant proximity to teeth; dental-contected implants; dentointegration;cementum formation on implant surface" in Chinese and English, respectively. RESULTS AND CONCLUSION: SST aims to maintain alveolar bone and gingival tissues by preserving partial tooth root with its periodontium-bundle bone intact, which is an alternative strategy for tissue preservation in esthetics zone.

Long-term Helicobacter pylori Infection Does Not Induce Tauopathy and Memory Impairment in SD Rats / 华中科技大学学报（医学）（英德文版）

Helicobacter pylori (H.pylori) infection is a recognized risk factor of dementia,while its role and mechanism in Alzheimer disease (AD) remained unclarified.Our previous study has identified that injection of soluble H.pylori filtrate could induce AD-like pathologic changes and cognitive impairment in SD rats.In the present study,we further explored the effect of long-term stomach colonization of H.pylori bacteria on the brains of SD rats.The results showed that H.pylori bacteria gavage induced an efficient colonization of H.pylori in the stomach after four weeks.However,there was no significant change of tau phosphorylation at Thr205 (pT205),Thr231 (pT231),Ser396 (pS396) and Ser404 (pS404) sites in the hippocampus and cerebral cortex.The H.pylori-infected rats also showed no cognitive impairment.These observations may result from inefficient release of bacterial pathogenic factors or the overall lack of host inflammatory responses.We conclude that SD rat with long-term H.pylori colonization in the stomach is not a suitable animal model for exploring the effects of H.pylori infection on brain function in human beings;administration of bacterial filtrates may better reveal the systemic pathologic changes induced by bacterial infection in animals which show a negative host response to bacterial colonization.

Three Dimensional Orbital Magnetic Resonance T2-Mapping in the Evaluation of Patients with Graves' Ophthalmopathy / 华中科技大学学报（医学）（英德文版）

The clinical application of orbital magnetic resonance (MR) T2-mapping imaging in detecting the disease activity of Graves' ophthalmopathy (GO),and the predictive values of therapy response to intravenous glucocorticoid (ivGC) were investigated.Approved by the local institutional review board (IRB),106 consecutive patients with GO were included in this prospective study.All subjects were divided into two groups according to the patients' clinical activity score (CAS):the CAS positive group (CAS ≥3) or the CAS negative group (CAS ＜3).T2 relaxation time of extraocular muscles (T2RT;ms) and the areas of four extra-ocular muscles (AEOMs;mm2) were measured by 3D T2-mapping MR sequence before and after methylprednisolone treatment,so as the CAS and some ophthalmic examinations including visual acuity,intra-ocular pressure,eyeball movement,diplopia and proptosis.In addition,24 healthy volunteers were recruited as the control group.The mean T2RT and AEOMs in CAS positive group were higher than those in CAS negative group.Both CAS positive and negative groups had significantly higher mean T2RT and AEOMs than the control group (P＜0.01).There was a positive correlation between T2RT and AEOMs values in GO patients,both of them had a positive correlation with CAS and the ophthalmic examinations.It was concluded that to evaluate the activity of GO,CAS was mostly related to inflammation symptoms of ocular surface,more than that,T2RT and AEOMs were also related to abnormal findings of the ophthalmic examinations including high ocular pressure,impaired eyeball movement,diplopia and proptosis.T2RT and AEOMs can reflex the inflammation state of ocular muscles better.CAS combined with 3D T2-mapping MR imaging could improve the sensitivity of detection of active GO so as the prediction and evaluation of the response to methylprednisolone treatment.

Short hairpin RNA targeting insulin-like growth factor binding protein-3 restores the bioavailability of insulin-like growth factor-1 in diabetic rats

ABSTRACT Purpose To investigate whether intracavernosal injection of short hairpin RNA for IGFBP-3 could improve erectile function in streptozotocin-induced diabetic rats. Materials and methods After 12 weeks of IGFBP-3 short hairpin RNA injection treatment, intracavernous pressure responses to electrical stimulation of cavernous nerves were evaluated. The expression of IGFBP-3 and IGF-1 at mRNA and protein levels were detected by quantitative real-time PCR analysis and Western blot, respectively. The concentration of cavernous cyclic guanosine monophosphate was detected by enzyme-linked immunosorbent assay. Results At 12 weeks after intracavernous administration of IGFBP-3 shRNA, the cavernosal pressure was significantly increased in response to the cavernous nerves stimulation compared to the diabetic group (P<0.05). Cavernous IGFBP-3 expression at both mRNA and protein levels was significantly inhibited. At the same time, cavernous IGF-1 expression was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Cavernous cyclic guanosine monophosphate concentration was significantly increased in the IGFBP-3 shRNA treatment group compared to the diabetic group (P<0.01). Conclusions Gene transfer of IGFBP-3 shRNA could improve erectile function via the restoration of cavernous IGF-1 bioavailability and an increase of cavernous cGMP concentration in the pathogenesis of erectile dysfunction in streptozotocin-induced diabetic rats.

Sex hormone levels may be an important factor influencing the gender dimorphism in prognosis of severe trauma patients / 中华急诊医学杂志

Objective To describe the association between age and the sex-based outcome difference and analyze the potential mechanism responsible for sex-based outcome difference in severe trauma patients.Methods A retrospective analysis derived from the Emergency Intensive Care Unit of the acute trauma center of the Shanghai Municipal Sixth People's Hospital during the 2010-2013 period was performed to identify sex-based outcome differences after severe blunt trauma.The study cohort of patients was then stratified by age:(1) 18 years ≤ age ＜45 years,(2) 45 years ≤ age ≤ 55 years,and (3) age ＞55 years.Crude and adjusted odds ratios (ORs) were calculated to evaluate the association between gender and the hospitalized mortality,both overall and subgroups according to age categories.Results A total of 987 severe trauma patients met our inclusion criteria were enrolled in this study.Crude mortality was higher in male severe trauma patients (male 9.1％ vs.female 5.0％,P ＜ 0.05).Multiple logistic regression revealed that females had a 79％ decrease in hospitalized mortality compared with males (OR =0.21,95％ CI:0.07-0.64,P =0.006).This difference was most distinct in patients with age ＜ 45 years,(OR =0.15,95％ CI:0.04-0.67,P =0.012).There is no significant difference between genders in mortality of trauma patients with 45 years ≤ age ≤ 55 years and age ＞ 55 years.Conclusions The present study revealed a statistical significant association between gender and mortality among severe blunt trauma patients,particularly in patients with age ＜ 45 years.These results may highlight the importance of sex hormones in outcomes of severe trauma.

Comparing the diagnostic values of circulating microRNAs and cardiac troponin T in patients with acute myocardial infarction

OBJECTIVE: Recent studies have shown that circulating microRNAs might be useful, novel biomarkers for the diagnosis of acute myocardial infarction. The aims of this study were to evaluate the expression of cardiac-specific miRNAs (miR-1, -133a, -208b, and -499) in patients with acute myocardial infarction and to compare the diagnostic values of these miRNAs with that of cardiac troponin T. METHODS: Sixty-seven plasma samples obtained from patients with acute myocardial infarction and 32 plasma specimens collected from healthy volunteers were analyzed in this study. The levels of cardiac-specific miRNAs (miR-1, -133a, -208b, and -499) were measured by quantitative reverse transcription-polymerase chain reaction, and the concentrations of plasma cardiac troponin T were measured using electrochemiluminescence-based methods and an Elecsys 2010 Immunoassay Analyzer. RESULTS: The levels of plasma miR-1, -133a, -208b, and -499 were significantly higher in acute myocardial infarction patients (all p<0.001) than in healthy volunteers. The expression of the cardiac-specific miRNAs in acute myocardial infarction patients decreased to close to the baseline levels at the time of hospital discharge (all p>0.05). There were no correlations between the levels of the four circulating miRNAs and the clinical characteristics of the study population (all p>0.05). Furthermore, receiver operating characteristic curve analyses showed that the four plasma miRNAs were not superior to cardiac troponin T for the diagnosis of acute myocardial infarction (all p>0.05). CONCLUSION: Our results demonstrate that circulating miR-1, -133a, -208b, and -499 may be useful biomarkers in acute myocardial infarction patients but that these miRNAs are not superior to cardiac troponin T for the diagnosis of acute myocardial infarction.

Clinical Effects of Intensive Insulin Therapy Treating Traumatic Shock Combined with Multiple Organ Dysfunction Syndrome / 华中科技大学学报（医学）（英德文版）

The therapeutic effects of intensive insulin therapy in treatment of traumatic shock combined with multiple organ dysfunction syndrome (MODS) were investigated.A total of 114 patients with traumatic shock combined with MODS were randomly divided into two groups:control group (n=56) treated with conventional therapy,and intensive insulin therapy group (n=58) treated with conventional therapy plus continuous insulin pumping to control the blood glucose level at range of 4.4-6.1 mmol/L.White blood cells (WBC) counts,prothrombin time (PT),serum creatinine (SCr),alanine aminotransferase (ALT),serum albumin and PaO2 were measured before and at the day 1,3,5,7 and 14 after treatment.The incidence of gastrointestinal dysfunction,the incidence of MODS,hospital stay and the mortality were also observed and compared.After intensive insulin therapy,the WBC counts,SCr,ALT and PT were significantly reduced (P＜0.05),but the level of serum albumin was significantly increased (P＜0.05) at the day 3,5,7 and 14.In the meantime,the PaO2 was significantly elevated at the day 3,5 and 7 (P＜0.01) after intensive insulin therapy.The incidence of gastrointestinal dysfunction,the incidence of MODS,the length of hospital stay and the mortality were markedly decreased (P＜0.01).The results suggest early treatment with intensive insulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

Evaluation of Atrial Septal Defect Using Real-time Three-dimensional Echocardiography:Comparison with Surgical Findings / 华中科技大学学报（医学）（英德文版）

The present study evaluated the application of three dimensional echocardigraphy (3DE) in the diagnosis of atrial septal defect (ASD) and the measurement of its size by 3DE and compared the size with surgical findings.Two-dimensional and real-time three dimensional echocardiography (RT3DE) was performed in 26 patients with atrial septal defect,and the echocardiographic data were compared with the surgical findings.Significant correlation was found between defect diameter by RT3DE and that measured during surgery (r=0.77,P<0.001).The defect area changed significantly during cardiac cycle.Percentage change in defect size during cardiac cycle ranged from 6%-70%.Our study showed that the size and morphology of atrial septal defect obtained with RT3DE correlate well with surgical findings.Therefore,RT3DE is a feasible and accurate non-invasive imaging tool for assessment of atrial septal size and dynamic changes.

Assessment of Myocardial Perfusion and Systolic Function in Patients with Coronary Artery Disease after Coronary Artery Bypass Surgery by Myocardial Contrast Echocardiography and Two-dimensional Strain Echocardiography / 华中科技大学学报（医学）（英德文版）

The clinically applied value of myocardial perfusion and systolic function in patients with coronary artery disease after coronary artery bypass surgery using real-time myocardial contrast echo-cardiography (RT-MCE) combined with two-dimensional strain echocardiography was assessed.Twenty patients underwent intravenous RT-MCE by intravenous injections of Sono Vue before and after coronary artery bypass surgery. Two-dimensional images were recorded from the left ventricular four-chamber view, two-chamber view and the apical view before, and two weeks and three months af-ter coronary artery bypass surgery, and the peak systolic longitudinal strain was measured. The results showed that myocardial perfusion was significantly increased after coronary artery bypass surgery in about 71.6% segments. In the group that myocardial perfusion was improved, the peak systolic longitu-dinal strain three months after bypass surgery was significantly higher than that before operation [(-15.78±5.91)% vs (-10.45±8.31)%, P<0.05]. However, the parameters did not change in the group without myocardial perfusion improvement [(-10.33±6.53)% vs (-9.41±6.09)%, P>0.05]. It was con-cluded that whether or not the improvement of myocardial perfusion can mirror the recovery trend of regional systolic function, two-dimensional strain echocardiography can observe dynamic change of re-gional systolic function. The combination of myocardial perfusion with two-dimensional strain echocar-diography can more accurately assess the curative effectiveness of coronary artery bypass surgery.

Autoantibodies to the high-affinity IgE receptor in patients with asthma.

Autoimmune diseases have been implicated as a cause of intrinsic asthma; however, there is little data on the role of autoimmunity in the pathogenesis of asthma. The purpose of this study was to investigate circulating autoantibodies against the high-affinity IgE receptor Fc(epsilon)RI in patients with asthma. Seventy-eight patients with asthma and 32 healthy individuals as control subjects were included. All subjects were tested with basophil histamine releasing assay and immunoblotting to assess for the potential presence of receptor Fc(epsilon)RI autoantibodies. Of the 78 asthma patients total subjects, 25 (32.1%) had a positive by basophil histamine releasing assay and 23 (29.5%) by immunoblotting. Both of them were significant higher than the positive rate, 9.4% (p < 0.05) and 9.4% (p < 0.05), respectively. Our data demonstrated that aberrant autoantibodies against the high-affinity IgE receptor Fc(epsilon)RI were found in some patients with asthma implies that the autoimmunity may be one factor in intrinsic asthma pathogenesis.

Induction of SiHa Cells Cells Apoptosis by Nanometer Realgar Suspension and Its Mechanism / 华中科技大学学报（医学）（英德文版）

The effects of nanometer realgar suspension on proliferation and apoptosis of human uterine cervix cancer cell line SiHa cells and oncogenic genes HPV16E6/E7 were investigated. A "micro-jet efflux" strategy was used for the preparation of nanometer realgar suspension. SiHa cells were treated with nanometer Realgar suspension in various concentrations (6.25,12.5,25 and 50mg/L) for different durations (12,24,48 and 72h). The inhibitive effect of nanometer realgar suspension on growth of SiHa cells was detected by MIT method. Special morphological changes of apoptosis were observed by transmission electron microscopy (TEM) and DNA fragments electrophoresis. The apoptotic rate was quantified by flow cytometry (FCM). The expression of HPV16E6/E7 mRNA and protein was assayed by RT-PCR and Western blot respectively. The results showed after being treated with 25-50mg/L nanometer realgar suspension for 48h, the survival rate of SiHa cells was decreased, and apoptotic rate markedly increased in a time- and concentration-dependent manner. TEM and DNA electrophoresis revealed the special morphological changes of apoptosis. The apoptotic rate of SiHa cells treated with nanometer realgar suspension was significantly higher than in the control group (P<0.01), and G0/G1 phase arrest appeared following treatment with nanometer realgar suspension in 25 and 50mg/L for 48h.RT-PCR and Western blot assay indicated that nanometer realgar suspension reduced the HPV16E6/E7 gene expression. Nanometer realgar suspension could inhibit the proliferation and induce apoptosis of SiHa cells. The mechanism may be related to the down-regulation of the HPV16E6/E7 gene expression.

Effect of dexamethasone on spontaneously apoptosis of polymorphonuclear neutrophils from postburn rabbits / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the effect of dexamethasone on spontaneously apoptosis, bcl-2, and neuclear facor kappa (NFkappaB) expressions of polymorphonuclear neutrophil (PMN) from postburn rabbits.</p><p><b>METHODS</b>PMN were isolated from 8 rabbits on 24 postburn hours and cultured with normal serum (NS), burn serum (BS), normal serum plus dexamethasone (ND), and burn serum plus dexamethasone (BD) for 24 hours, respectively. The quantification of apoptosis was analyzed by acridine orange + ethidium bromide fluorescent staining and flow cytometry , and the contents of bcl-2 and NFkappaB protein detected by immunohistochemical method.</p><p><b>RESULTS</b>In the BS group, the percentage of apoptotic PMN decreased (compared with NS group, 6.18 +/- 0.96 vs. 21.77 +/- 2.32, P<0.05), and the contents of bcl-2 and NFkappaB protein increased (compared with NS group, 83.27 +/- 5.45 vs. 49.95 +/- 2.67, P<0.05). When compared with BS group, the apoptotic percentage of BD group increased (12.67 +/- 0.71 vs. 6.18 +/- 0.96, P<0.05), and the content of NFkappaB protein reduced (0. 1031 +/- 0.0154 vs. 0.1802 +/- 0.0130, P<0.05), but no significant difference between ND and NS group was found.</p><p><b>CONCLUSION</b>Dexamethasone decreases the inhibition of PMN apoptosis by bumn serum, which may be associated with the down-regulation of NFKB expression.</p>

Construction and application of a yeast expression system for thymosin a1

We want to construct a yeast expression system for thymosin a1 (Ta1) to make the orally administered Ta1 preparation possible. The whole Ta1 DNA fragment was obtained by PCR. After being digested with restriction enzymes, it was cloned into pYES2 vector. Sequencing was performed to identify the recombinant. The sequence of Ta1 in recombinant coincided with the original one reported in Genbank. When pYES2-Ta1 plasmid was transformed into yeast, galactose instead of glucose was used to induce Ta1 expression. Western blot was performed to identify the quality of the expressed Ta1. Dried yeast containing pYEST2-Ta1 was fed to Balb/c mice whose immunities were inhibited by cyclophosphamide in advance. Synthesized Ta1 peptide was used as positive control and empty yeast was used as negative control. Compared with the negative control group, both dried yeast containing pYEST2-Ta1 and synthesized Ta1 peptide can significantly increase the CD8+ level (22.74±1.09 and 18.77±4.72 vs 7.49±2.14, p<0.01), while both of them had little effect on the CD4+ lymphocytes (61.86±6.94 and 65.91±4.78 vs 57.93±10.40, p>0.05). We concluded that a high effective yeast expression system for Ta1 was constructed successfully and the Ta1 protein expressed by this system can improve CD8+ level in immune inhibited mice.

High expression of the foot-and-mouth disease's structural protein P1 in Escherichia coli and analysis of its biology activity / 生物工程学报

Foot-and-mouth disease virus (FMDV) is the aetiological angent of a highly contagious viral disease. The complete gene encoding the structural protein of FMDV (P1) was subcloned into expression vector pGEX-KG, resulting in the fusion expression plasmid pKG-P1. After transformed into E. coli BL21(DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-P1 fusion protein was expressed in high level. The molecular weight of the fusion protein wa 110kD and the expressed products were soluble. Western-blotting was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV. The fusion proteins were further purified by GST purification kit and an indirect ELISA (P1-ELISA) based on the purified proteins was developed. Comparison between P1-ELISA and the standard indirect haemagglutinin assay showed the two methods had 87 per cent agreement by detecting 864 serum samples, indicating the purified P1 protein was specific as the antigen of indirect P1-ELISA.

Expression of GP5-M fusion protein of porcine reproductive and respiratory syndrone virus (PRRSV) and establishment of ELISA diagnose based on the recombinant fusion protein / 生物工程学报

The cDNA fragment encoding the truncated GP5 and the full-length M protein of Porcine Reproductive and Respiratory Syndrone Virus (PRRSV) were orderly fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pKG-56. After transformed into E. coli BL21 (DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-GP5-M fusion protein was expressed in high level. Western-blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against PRRSV. The fusion protein was further purified and used as an antigen to establish a novel PRRSV ELISA diagnose assay (P56-ELISA). Comparison between P56-ELISA and the abroad kit IDEXX-ELISA showed the two methods had 94.1 percent agreement by detecting 205 serum samples, indicating that the indirect P56-ELISA was specific and sensitive. The correlation between virus neutralization antibody of the infected pigs (not convalescent pigs) and antibody response to the fusion protein GP5-M was further studied. The regression function analysis suggested that there was no significant correlation between ELISA antibody response (OD630 nm) to the fusion protein GP5-M in clinical serum and their specific neutralizing titers.

Immunogenicity of DNA vaccine expressing GP5 of porcine reproductive and respiratory syndrome virus fused with VP22 of bovine herpesvirus 1 / 生物工程学报

To enhance the immuogenicity of DNA vaccines expressing the GP5 protein of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the tegument protein VP22 (encoded by VP22 gene) of Bovine Herpesvirus 1 (BHV-1), which has been demonstrated to exhibit the unusual protein transduction property, was fused to N-terminus of GP5 of DNA vaccine construct pCI-ORF5M, resulting in pCI-VP22-ORF5M expressing VP22-GP5 fusion protein. The expression of VP22-GP5 fusion protein was confirmed by both indirect immunofluorescence assay (IFA) and Western blot. To investigate its immunogenicity, BALB/c mice were immunized with the fusion expression plasmid pCI-VP22-ORF5M and non-fusion expression plasmid pCI-ORF5M, respectively. The GP5-specific ELISA antibodies, neutralizing antibodies and lymphocyte proliferative responses were evaluated at various time points after primary immunization. The results showed that GP5-specific ELISA antibodies, neutralizing antibodies, and lymphocyte proliferative responses induced by DNA vaccine pCI-VP22-ORF5M were higher significantly than those of DNA vaccine pCI-ORF5M, indicating that fusion expression with BHV-1 VP22 significantly enhances the immuogenicity of DNA vaccine expressing the PRRSV GP5 protein, and that this strategy may also be useful to develop more efficient DNA vaccines against other pathogens.