Research advances in vaccines against coxsackievirus group B / 中华微生物学和免疫学杂志

Coxsackievirus group B (CVB) belongs to human enterovirus and contains six serotypes. CVB infection in humans usually causes mild symptoms, but can lead to myocarditis, pancreatitis, hand, foot and mouth disease and central nervous system diseases, which is extremely harmful to infants and young children. There are outbreaks of CVB in many places at home and abroad. The epidemic patterns of different serotypes are distinct and complicated. Currently, there are no effective drugs or means to treat or prevent CVB infection. Vaccine is expected to be a cost-effective way to address CVB-related public health concerns. Although there is no CVB vaccine on the market, many institutions are conducting preclinical studies of various types of CVB vaccines, including nucleic acid vaccines, viral vector vaccines, recombinant protein vaccines, live-attenuated vaccines and inactivated vaccines. This article reviewed the advances in the research of CVB vaccines.

Clinical efficacy of different treatments and their impacts on the quality of life of octogenarians with coronary artery disease / 中华医学杂志(英文版)

BACKGROUND@#Coronary artery disease (CAD) in octogenarians (age of ≥80 years) has a high risk of mortality and high medical expenses. Research shows that the prevalence of CAD is higher among octogenarians than that among younger people, but few such patients undergo percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG). This study aimed to evaluate different treatments with respect to their clinical effects and impacts on quality of life of octogenarians with CAD.@*METHODS@#Data of 519 octogenarians with CAD consecutively treated at Beijing Anzhen Hospital, Capital Medical University (Beijing, China) from January 2010 to January 2016 were collected in this study. The patients were categorized into three groups based on the treatments they received: the PCI group (n = 292), CABG group (n = 110), and medical treatment group (n = 117). The followings were recorded during follow-up: clinical data, death (all-cause and cardiovascular-related), re-hospitalization time, Seattle Angina Questionnaire (SAQ) score, and occurrence of hemorrhagic events (cerebral bleeding, gastrointestinal bleeding, and dermal ecchymosis).@*RESULTS@#The median follow-up duration was 25.0 (25th, 75th percentile: 17.0, 55.5) months among 417 patients. The all-cause death rates (28.2% vs. 12.0% and 14.6%, respectively) and cardiovascular-related death rates (15.4% vs. 3.8% and 6.4%, respectively) were significantly higher in the medical treatment group than those in the PCI group and CABG group (all P < 0.05). The re-hospitalization rate for cardiovascular events was significantly lower in the CABG group than those in the PCI group and medical treatment group (3.8% vs. 12.8% and 14.9%, respectively) (χ = 8.238, P = 0.018). The SAQ scores of physical limitation, angina frequency, treatment satisfaction, and disease perception were significantly higher in the PCI group and CABG group than those in the medical treatment group (all P < 0.05). No significant difference in the angina stability score was observed among the three groups (F = 3.179, P = 0.204).@*CONCLUSION@#PCI and CABG result in reduced mortality and better quality of life in octogenarians with CAD.

Identification of three novel SRD5A2 mutations in Chinese patients with 5α-reductase 2 deficiency / 亚洲男科学杂志(英文版)

In this study, we investigated the genetics, clinical features, and therapeutic approach of 14 patients with 5α-reductase deficiency in China. Genotyping analysis was performed by direct sequencing of PCR products of the steroid 5α-reductase type 2 gene (SRD5A2). The 5α-reductase activities of three novel mutations were investigated by mutagenesis and an in vitro transfection assay. Most patients presented with a microphallus, variable degrees of hypospadias, and cryptorchidism. Eight of 14 patients (57.1%) were initially reared as females and changed their social gender from female to male after puberty. Nine mutations were identified in the 14 patients. p.G203S, p.Q6X, and p.R227Q were the most prevalent mutations. Three mutations (p.K35N, p.H162P, and p.Y136X) have not been reported previously. The nonsense mutation p.Y136X abolished enzymatic activity, whereas p.K35N and p.H162P retained partial enzymatic activity. Topical administration of dihydrotestosterone during infancy or early childhood combined with hypospadia repair surgery had good therapeutic results. In conclusion, we expand the mutation profile of SRD5A2 in the Chinese population. A rational clinical approach to this disorder requires early and accurate diagnosis, especially genetic diagnosis.

Clinical efficacy of different treatments and their impacts on the quality of life of octogenarians with coronary artery disease / 中华医学杂志(英文版)

Background@#Coronary artery disease (CAD) in octogenarians (age of ≥80 years) has a high risk of mortality and high medical expenses. Research shows that the prevalence of CAD is higher among octogenarians than that among younger people, but few such patients undergo percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG). This study aimed to evaluate different treatments with respect to their clinical effects and impacts on quality of life of octogenarians with CAD.@*Methods@#Data of 519 octogenarians with CAD consecutively treated at Beijing Anzhen Hospital, Capital Medical University (Beijing, China) from January 2010 to January 2016 were collected in this study. The patients were categorized into three groups based on the treatments they received: the PCI group (n = 292), CABG group (n = 110), and medical treatment group (n = 117). The followings were recorded during follow-up: clinical data, death (all-cause and cardiovascular-related), re-hospitalization time, Seattle Angina Questionnaire (SAQ) score, and occurrence of hemorrhagic events (cerebral bleeding, gastrointestinal bleeding, and dermal ecchymosis).@*Results@#The median follow-up duration was 25.0 (25th, 75th percentile: 17.0, 55.5) months among 417 patients. The all-cause death rates (28.2% vs. 12.0% and 14.6%, respectively) and cardiovascular-related death rates (15.4% vs. 3.8% and 6.4%, respectively) were significantly higher in the medical treatment group than those in the PCI group and CABG group (all P < 0.05). The re-hospitalization rate for cardiovascular events was significantly lower in the CABG group than those in the PCI group and medical treatment group (3.8% vs. 12.8% and 14.9%, respectively) (χ2 = 8.238, P = 0.018). The SAQ scores of physical limitation, angina frequency, treatment satisfaction, and disease perception were significantly higher in the PCI group and CABG group than those in the medical treatment group (all P < 0.05). No significant difference in the angina stability score was observed among the three groups (F = 3.179, P = 0.204).@*Conclusion@#PCI and CABG result in reduced mortality and better quality of life in octogenarians with CAD.

Progress in clinical research of varicella vaccines / 中华微生物学和免疫学杂志

Varicella(chickenpox)is an acute infectious disease with high incidence in children. It is mainly transmitted through the airborne route and vaccination is the best measure for the prevention and control. Data from post-marketing studies show the effectiveness of varicella vaccines is 80%-85%, and two-dose regimen is significantly more effective than one-dose. After inclusion of varicella vaccines into the national immunization programme, there has been a clear decrease in varicella morbidity. Despite the lack of direct evidence, there remains the risk that varicella-zoster virus might latent in the dorsal route ganglia after vaccination. Therefore, more safe and effective novel varicella vaccines are under development. This paper reviewed the progress in varicella vaccine development and their long-term efficacy and safety.

Progress in clinical research of varicella vaccines / 中华微生物学和免疫学杂志

Varicella( chickenpox) is an acute infectious disease with high incidence in children. It is mainly transmitted through the airborne route and vaccination is the best measure for the prevention and control. Data from post-marketing studies show the effectiveness of varicella vaccines is 80％-85％, and two-dose regimen is significantly more effective than one-dose. After inclusion of varicella vaccines into the na-tional immunization programme, there has been a clear decrease in varicella morbidity. Despite the lack of direct evidence, there remains the risk that varicella-zoster virus might latent in the dorsal route ganglia after vaccination. Therefore, more safe and effective novel varicella vaccines are under development. This paper reviewed the progress in varicella vaccine development and their long-term efficacy and safety.

Activities of polysaccharides in combination with aluminum adjuvant in varicella-zoster subunit vaccines / 中华微生物学和免疫学杂志

Objective To evaluate the immunostimulatory effects of polysaccharides, including Lycium barbarum polysaccharides ( LBP) , Angelica polysaccharides ( AP) , Licorice polysaccharides ( LP) and Inulin polysaccharides ( IP) , used alone or in combination with aluminum adjuvant on the recombinant protein of varicella-zoster virus (VZV) glycoprotein (gE) as an immunogen in mice. Methods BALB/c mice were randomly divided into 11 groups with five in each group. Intramuscular injection was given at Days 0 and 14. Serum samples were collected at weeks 0, 2, 3, 4 and detected for total anti-gE IgG titers and an-tibody subtypes by indirect ELISA. The mice were sacrificed at week 4 to collect spleen lymphocytes aseptic-ally. CCK-8 method was used to evaluate the proliferation of spleen lymphocytes. The levels of IFN-γ and IL-4 were measured by ELISA. The percentages of CD3+CD4+ and CD3+CD8+ T cell subsets were deter-mined by flow cytometry. Results The four plant-derived polysaccharides in combination with aluminum adjuvant showed good adjuvant activities. LP combined with aluminum adjuvant effectively increased the titer of IgG2, promote the proliferation of splenocytes and enhanced the secretion of IFN-γ and IL-4. Further-more, it also induced CD3+CD4+ and CD3+CD8+ T cell proliferation in vivo. Conclusions The four plant-derived polysaccharides in combination with aluminum adjuvant could all enhance antibody production. LP combined with aluminum adjuvant could induce cell-mediated immune responses, suggesting that it might be a promising adjuvant for varicella-zoster subunit vaccines.

Unfractionated Heparin with Sequential Enoxaparin in Patients with Complex Coronary Artery Lesions during Percutaneous Coronary Intervention / 中华医学杂志(英文版)

<p><b>Background</b>Unfractionated heparin (UFH), despite its limitations, has been used as the primary anticoagulant alternative during the percutaneous coronary intervention (PCI). Some studies indicated that intravenous enoxaparin could be an effective and safe option. Our team used enoxaparin alone at one time according to the guidelines (Class IIA) and found a little catheter thrombosis during PCI. We recommend a new anticoagulation strategy using enoxaparin in combination with UFH. Enoxaparin has a more predictable anticoagulant response with no need of repeatedly monitoring anticoagulation during PCI. This retrospective study aimed to evaluate the efficacy and safety of using enoxaparin in combination with UFH in PCI patients with complex coronary artery disease.</p><p><b>Methods</b>Between January 2015 and April 2017, 600 PCI patients who received intravenous UFH at an initial dose of 3000 U plus intravenous enoxaparin at a dose of 0.75 mg/kg (observation group) and 600 PCI patients who received UFH at a dose of 100 U/kg (control group) were consecutively included in this retrospective study. The endpoints were postoperative 48-h thrombolysis in myocardial infarction (TIMI) bleeding and transfusion and 30-day and 1-year major adverse cardio-cerebrovascular events (MACCE).</p><p><b>Results</b>Baseline clinical, angiographic, and procedural characteristics were similar between groups, except there was less stent implantation per patient in the observation group (2.13 vs. 2.25 in the control group, P = 0.002). TIMI bleeding (3.3% vs. 4.7%) showed no significant difference between the observation group and control group. During the 30-day follow-up, the rate of MACCE was 0.9% in the observation group and 1.5% in the control group. There was no significant difference in the rates of MACCE, death, myocardial infarction, target vessel revascularization, cerebrovascular event, and angina within 30 days and 1 year after PCI between groups as well as in the subgroup analysis of transfemoral approach.</p><p><b>Conclusions</b>UFH with sequential enoxaparin has similar anticoagulant effect and safety as UFH in PCI of complex coronary artery disease.</p>

Effects of recent upper respiratory-tract infections on incidence of the perioperative respiratory adverse events in children: a prospective cohort study / 北京大学学报(医学版)

Objective:To investigate the effects of the recent upper respiratory tract infections (URI) on the incidence of perioperative respiratory adverse events in children scheduled to undergo general anesthesia and elective surgery.Methods:In the study,232 children undergoing general anesthesia with laryngeal mask airway (LMA) for elective ophthalmic surgeries at Peking University First Hospital,Beijing,China,from Nov.1,2015 to May 10,2016 were enrolled.On the day of the surgery,the parents of the children were preoperatively asked to fill out a questionnaire regarding the baseline characteristics and medical history of the children,including gender,age,height,weight,history of URI within the last 2 weeks before anesthesia,history of premature,long-term passive smoking exposure,habitual sleep snoring,and history of asthma.In addition,all adverse respiratory events throughout the perioperative periods (oxygen desaturation,cough,copious secretions,laryngospasm and bronchospasm) as well as perioperative variables (number of attempts to insert the LMA successfully,anesthesia duration and so on) were recorded.Multivariate Logistic regression analysis was applied to identify independent risk factors of perioperative respiratory adverse events.Results:Among the 232 children included in the study,28.0％ (65/232) presented with a history of a recent URI within the last 2 weeks before anesthesia.The presence of the recent URI increased the incidence of oxygen desaturation (23.1％ vs.12.0％,P =0.034),copious secretions (15.4％ vs.6.6％,P =0.036) and any of all the adverse respiratory events (32.3％ vs.18.6％,P =0.024).Multivariate Logistic regression analysis identified two independent risk factors of perioperative adverse respiratory events:a history of URI within the last 2 weeks before general anesthesia (OR =2.021,95％ CI:1.023-3.994,P =0.043) and habitual sleep snoring (OR =3.660,95％ CI:1.517-8.832,P =0.004).Conclusion:A history of a recent URI within 2 weeks before general anesthesia was associated with a higher incidence of oxygen desaturation,copious secretions and the overall respiratory adverse events.For the children with recent URI,we recommend the general anesthesia and elective surgery should be postponed for at least 2 weeks after the URI.

Preparation of monoclonal antibodies against 3D protein of EV71 based on HBc particles as expression vector / 中国免疫学杂志

Objective:To prepare and preliminarily identify the monoclonal antibodies(mAbs) specifically against 3D protein of Enterovirus 71(EV71),using bioinformatics to predict the epitopes of 3D,with HBc protein as a carrier.Methods: Artificial screening of 3D protein epitope sequences by bioinformatic method,inserted into the major immunodominant region(MIR) area of Hepatitis B virus core protein(HBc),to construct the recombinant protein.BALB/c mice were immunized with the recombinant virus like particles(VLPs),to prepare the mAbs against 3D protein of EV71.Affinity chromatography technology was used to purify the mAb.The indirect ELISA,ELISPOT,immunofluorescence and immunohistochemistry staining methods were used to identify the characteristic of the mAb.Results: We displayed 3D(aa34-43),3D(aa61-76) and 3D(aa151-164) epitopes by constructing fusion protein using HBc VLPs as a vector,after hybridization,one positive hybridoma cell line(3E1) was selected by ELISA.The isotype of 3E1 was IgG2a.The results of immunofluorescence and immunohistochemistry staining assay showed that the mAb 3E1 could specifically recognize EV71.Conclusion: The prepared mAb 3E1 can specifically recognizes the EV71,which laid the foundation for the detection of virus and further study on 3D protein,and verified the bioinformatics technology combined with HBc carrier displaying peptides could prepare mAb quickly and efficiently.

Relationship between peripheral neuropathy and retinal nerve fiber layer thickness in elderly patients with type 2 diabetes / 中华老年医学杂志

Objective To investigate the relationship between retinal nerve fiber layer thickness and peripheral neuropathy in elderly patients with type 2 diabetes.Methods Clinical data of elderly patients with T2DM in Endocrinology Department in Beijing Hospital were retrospectively collected.Global and sectorial retinal nerve fiber layer(RNFL)thicknesses were measured by using optical coherence tomography(OCT),and never conduction velocity measurements were performed in all patients.Diabetic peripheral neuropathy was diagnosed by the criteria in diabetic neuropathies update(2010,American Diabetes Association).RNFL thickness was compared between diabetic nonperipheral neuropathy group (control group,n =30)and diabetic peripheral neuropathy group (DPN group,n=17)and between different sub-groups.Results The RNFL thicknesses of temporal,nasal,superior and inferior visual fields and the mean RNFL thickness were less in DPN group than in control group,among which there were significant differences in the RNFL thicknesses of superior,inferior visual fields and the mean RNFL thickness [(107.7±27.4)μm vs.(128.1±17.3)μm,(112.9 ±20.8)μm vs.(130.8±21.8)μm,(88.2±15.5)μm vs.(100.5± 11.3)μm,F=7.446,7.468,7.988,respectively,P=0.009,0.009 and 0.007].RNFL thickness was decreased along with the aggravation of DPN from the control group,the subclinical DPN group to the DPN group successively(all P＜ 0.05).Conclusions Retinal nerve fiber layer thickness is associated with diabetic peripheral neuropathy in elderly patients with type 2 diabetes,and the relationship is more significant in patients with serious DPN.

Effects of free fatty acids on catalytic activity of P450c17 in cultured NCI-H295R cells of adrenal gland / 中华内分泌代谢杂志

[Summary] The activities of 17α-hydroxylase and 17,20 lyase of cytochrome P450c17 were evaluated by ELISA in NCI-H295R cells after treatment with palmitate and oleate. The results showed that 0. 75 mmol/L plamitate did not influence the activity of 17α-hydroxylase, but increased the activity of 17, 20 lyase by 74. 3% ( P<0. 05). Oleate at the same concentration did not change the activity of 17,20 lyase. There were no significant changes in the protein expressions of P450c17, P450 oxidoreductase, and cytochrome b5 after treatment with palmitate and oleate. However,reactive oxygen species in cells were elevated by palmitate. The results suggest that exposure to palmitate may increase androgen production by inducing 17, 20 lyase activity of P450c17 in NCI-H295R cells, which is related with oxidative stress-mediated post-translational regulation of the enzyme.

Protein kinase A-mediated cardioprotection of Tongxinluo relates to the inhibition of myocardial inflammation, apoptosis, and edema in reperfused swine hearts / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Our previous studies have demonstrated that Tongxinluo (TXL), a traditional Chinese medicine, can protect hearts against no-reflow and reperfusion injury in a protein kinase A (PKA)-dependent manner. The present study was to investigate whether the PKA-mediated cardioprotection of TXL against no-reflow and reperfusion injury relates to the inhibition of myocardial inflammation, edema, and apoptosis.</p><p><b>METHODS</b>In a 90-minute ischemia and 3-hour reperfusion model, minipigs were randomly assigned to sham, control, TXL (0.05 g/kg, gavaged one hour prior to ischemia), and TXL + H-89 (a PKA inhibitor, intravenously and continuously infused at 1.0 µg/kg per minute) groups. Myocardial no-reflow, necrosis, edema, and apoptosis were determined by pathological and histological studies. Myocardial activity of PKA and myeloperoxidase was measured by colorimetric method. The expression of PKA, phosphorylated cAMP response element-binding protein (p-CREB) (Ser(133)), tumor necrosis factor α (TNF-α), P-selectin, apoptotic proteins, and aquaporins was detected by Western blotting analysis.</p><p><b>RESULTS</b>TXL decreased the no-reflow area by 37.4% and reduced the infarct size by 27.0% (P < 0.05). TXL pretreatment increased the PKA activity and the expression of Ser(133) p-CREB in the reflow and no-reflow myocardium (P < 0.05). TXL inhibited the ischemia-reperfusion-induced elevation of myeloperoxidase activities and the expression of TNF-α and P-selectin, reduced myocardial edema in the left ventricle and the reflow and no-reflow areas and the expression of aquaporin-4, -8, and -9, and decreased myocytes apoptosis by regulation of apoptotic protein expression in the reflow and no-reflow myocardium. However, addition of the PKA inhibitor H-89 counteracted these beneficial effects of TXL.</p><p><b>CONCLUSION</b>PKA-mediated cardioprotection of TXL against no-reflow and reperfusion injury relates to the inhibition of myocardial inflammation, edema, and apoptosis in the reflow and no-reflow myocardium.</p>

Severe coronary artery disease in Chinese patients with abdominal aortic aneurysm: prevalence and impact on operative mortality / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Little is known about the prognosis of coronary artery disease (CAD) in Chinese patients with abdominal aortic aneurysm (AAA). The aim of this study was to evaluate the predictors of in-hospital all-cause mortality of severe CAD in Chinese patients who were hospitalized for AAAs.</p><p><b>METHODS</b>From January 2003 to August 2009, 368 patients were operated on for AAAs. The clinical characteristics were retrospectively collected. The primary outcome was the in-hospital all-cause mortality. The clinical risk factors were subjected to a multivariate analysis to determine the predictors of in-hospital all-cause mortality.</p><p><b>RESULTS</b>During their hospitalization, 23% (85/368) of the patients underwent coronary angiography, which revealed significant lesions in 93% (79/85) of the patients. In 25 cases, coronary artery bypass grafting (CABG) was performed before the AAA repair and in 16 cases of percutaneous coronary intervention (PCI) was performed. Ten patients with AAA alone died before discharge, and eight patients diagnosed with AAA combined with CAD died. There was no statistical difference in the postoperative death between the two groups. The logistic analysis showed that age > 70 years and CAD (vessels ≥ 2) were the significant factors in predicting the adverse clinical outcome.</p><p><b>CONCLUSIONS</b>The prevalence of severe CAD in Chinese patients with AAAs seemed lower than those that were reported. Myocardial evaluation and subsequent revascularization before AAA surgery could improve the clinical outcome for these patients who have severe CAD.</p>

Tacrolimus-related hypertrophic cardiomyopathy in an adult cardiac transplant patient / 中华医学杂志(英文版)

Left ventricular hypertrophy associated with the use of tacrolimus is a rare complication of solid organ transplantation in adult recipients. We present a cardiac transplant recipient who developed severe concentric left ventricular hypertrophy with congestive heart failure related to myocardial hypertrophy on tacrolimus. Hypertrophy improved when the drug was discontinued and replaced with sirolimus.

Ischemic preconditioning attenuates myocardial no-reflow and reperfusion injury after revascularization of acute myocardial infarction by reducing myocardial edema via the protein kinase A pathway / 中华心血管病杂志

<p><b>OBJECTIVE</b>Myocardial edema plays an important role in the development of myocardial no-reflow and reperfusion injury after the revascularization of acute myocardial infarction (AMI). The present study investigated whether the effect of ischemic preconditioning (IPC) against myocardial no-reflow and reperfusion injury was related to the reduction of myocardial edema through the protein kinase A (PKA) pathway.</p><p><b>METHODS</b>Twenty-four minipigs were randomized into sham, AMI, IPC, and IPC + H-89 (PKA inhibitor, 1.0 µg · kg(-1) · min(-1)) groups. The area of no-reflow (ANR), area of necrosis (AN), and water content in left ventricle and ischemic-myocardium and non-ischemic area were determined by pathological studies. Microvascular permeability was determined by FITC-labeled dextran staining. Cardiomyocyte cross-sectional area (CSA) and mitochondria cross-sectional area (MSA) were evaluated by histological analysis. Myocardial expression of aquaporins (AQPs) was detected by Western blot.</p><p><b>RESULTS</b>Compared with the MI group, the sizes of no-reflow and infarct were reduced by 31.9% and 46.6% in the IPC group (all P < 0.01), water content was decreased by 5.7% and 4.6% in the reflow and no-reflow myocardium of the IPC group (all P < 0.05), microvascular permeability and cardiomyocytes swelling in the reflow area were inhibited by 29.8% and 21.3% in the IPC group (all P < 0.01), mitochondrial water accumulation in the reflow and no-reflow areas of the IPC group were suppressed by 45.5% and 34.8% respectively (all P < 0.01), and the expression of aquaporin-4, -8, and -9 in the reflow and no-reflow myocardium were blocked in the IPC group. However, these beneficial effects of IPC were partially abolished in the IPC + H-89 group.</p><p><b>CONCLUSIONS</b>The cardioprotective effects of IPC against no-reflow and reperfusion injury is partly related to the reduction of myocardial edema by inhibition of microvascular permeability and aquaporins up-regulation via PKA pathway.</p>

Effects of mechanical ventilation on cell apoptosis in lung tissue of rat with acute lung injury / 中华急诊医学杂志

Objective To investigate the effects of different tidal volumes and positive end expiratory pressures on cell apoptosis in lung tissue of rats with acute lung injury.Methods Forty healthy male Sprague-Dawley rats were randomly(random number)divided into five groups,namely low tidal volume group(LV,VT 8 mL/kg),high tidal volume group(HV,VT 30 mL/kg),low tidal volume group with PEEP 2cmH2O(LV2P,VT 8 mL/kg,PEEP 2 cmH2O),low tidal volume group and PEEP 5cmH2O (LV5P,VT 8 mL/kg,PEEP 5 cmH2O)and low tidal volume group and PEEP 8 cmH2O(LV8P,VT 8 mL/kg,PEEP 8 cmH2O).After intravenous administration of oleic acid(OA,0.1 mL/kg),the rat model of acute lung injury was made.Mechanical ventilation was employed in rats of the experiment groups.Rats were sacrificed and their whole lungs were taken after mechanical ventilation for 2 hours.The transferase d-UTP end labeling assay(TUNEL)was used to define the extent and distribution of apoptotic cells in bronchus and lung tissues.The level of caspase-3 protein was determined by immunohistochemistry.Results The apoptotic cells on both alveolar septum and bronchial epithelium obviously increased with high level of caaspase-3 protein in HV group.The number of apoptotic cells obviously decreased with decrease in caspase-3 protein after PEEP ventilation.These changes were more significant in LV5P than those in other groups(P ＜ 0.01).Conclusions The mechanical ventilation with low tidal volume and PEEP produces protective effects on lung from injury.The cell apoptosis plays an important role in the course of VILI.

Construction and in vitro antiviral activity of highly effective artificial miRNA targeting to HIV-1 pol / 生物工程学报

RNA interference (RNAi) has exhibited huge potentials on anti-HIV-1 therapy research. The obtainment of RNAi element targeting to HIV-1 highly effectively and specifically was crucial for relevant research. Recent reports had described that microRNAs (miRNAs) posses more characteristics of inhibition and expression mechanisms than small interfering RNAs (siRNAs). In this study we explored the construction of artificial miRNA targeting to HIV-1 effectively and specifically. Sixteen siRNAs sequences were selected based on the conserved regions in the HIV-1 pol gene. ShRNA expression vectors were co-transfected with HIV-1 clone pNL4-3 to evaluate the abilities of siRNAs to inhibit HIV-1 expression. The pol1026 sequence was selected from candidates. The target sequence in the stem-loop structure of the well-characterized native miR-30a was replaced with pol1026 sequences, and the artificial miRNA expression vectors were co-transfected with the HIV-1 clone pNL4-3, results showed that HIV-1 can be effectively inhibited by miR-1026E. Target specificity of miR-1026E was confirmed by co-transfection assay with reporter plasmids containing different target sequences. The miR-1026E expression element was then inserted into Lentivirus which was used as a vector to transduce the MT-4 cells, MT-4-miR1026E expressing miR-1026E stably was cloned from transduced cells. The MT-4-miR1026E cell effectively inhibited HIV-1 replication in vitro. And the intracellular miR-181 and miR-16 expression levels and statl mRNA levels were not affected by the expression of miR-1026E in MT-4-miR1026E cells. miR-1026E is a promising candidate for future research.

Construction of highly effective artifical miRNA targeted to HIV-1 vif and the lentiviral-mediated antiviral research in vitro / 病毒学报

Effective and specific RNA interference (RNAi) elements are essential for the RNAi-based anti-HIV-1 research which has achieved extensive application. vif37 targeted to HIV-1 vi f is a highly effective and conserved RNAi target obtained from the previous study on screening. In this study, we explored the construction of artificial miRNAs to induce RNAi targeted to vif37, which had advantages on inhibition efficiency and flexibility of promoter selection. Three artificial miRNA targeted to vif37 were constructed by walking method using native miR-155 as a backbone and expressed by RNA polymerase II promoter. Then, expression vectors of artificial miRNA were co-transfected with HIV-1 infectious clone pNL4-3 to score its inhibition ability and showed that only miR-vif37 had the significant inhibition efficiency similar to shRNA-vif37. Subsequently, co-transfections with luciferase reporter plasmids into which different target sequences were inserted proved the specificity of miR-vif37 H. The replication of HIV-1 was inhibited in MT-4-miR37 H cells which could express miR-vif37 H stably and were cloned from MT-4 cells transducted with recombinant lentiviral vectors containing the miR-vif37 H expression element. Real-time RT-PCR revealed that miR-vif37 H had much lower expression level than shRNA-vif37. Results also showed that intracellular miR-181 and miR-16 expression levels and stat1 mRNA levels were not effected by the expression of miR-vif37 H in MT-4-miR37 H cells. We conclude miR-vif37 is a specific and highly effective artificial miRNA which will promote the further application of vif37 target.

Tetracysteine as a reporter for gene therapy / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the feasibility of using tetracysteine (TC) reporter in gene therapy.</p><p><b>METHODS</b>Effects of TC reporter and conventional reporter genes encoding green fluorescence protein (GFP) and luciferase (Luc) on expression and function of the therapeutic gene MGMT(P140K) were compared. Cytotoxicity and drug resistance were studied by Western blot. TC reporter used in therapy was analyzed by flow cytometry (FCM).</p><p><b>RESULTS</b>The TC reporter had no toxicity to cells and neither affected the expression or activity of therapeutic gene as compared to GFP and Luc. TC could be used in blood sample detection.</p><p><b>CONCLUSION</b>TC is a new kind of reporter gene for lentiviral vector in future gene therapy.</p>