Comparison of clinical efficacy of intravitreal injection of Conbercept and Aflibercept in the treatment of wet age-related macular degeneration / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To compare the clinical efficacy of conbercept and aflibercept in the treatment of wet age-related macular degeneration(wARMD)based on 4 consecutive intravitreal injections.METHODS: The clinical data of 108 patients(108 eyes)who were diagnosed as wARMD and treated with intravitreal injection at our hospital from January 2019 to January 2021 were retrospectively analyzed. They were divided into conbercept group(54 cases, 54 eyes)and aflibercept group(54 cases, 54 eyes)according to the different injectable drugs. All patients received intravitreal injection once a month, with four consecutive injections. Follow up for 12mo to observe best corrected visual acuity(BCVA), central macular thickness(CMT), complications and recurrence before and after injection.RESULTS: BCVA and CMT of patients in the two groups at 1, 2, 5 and 8mo after injection had no between-group differences(P>0.05), but both were significantly improved compared with those before injection(P<0.05). By the end of follow-up, conjunctival hemorrhage occurred in 2 eyes of the conbercept group at the early stage, and increased intraocular pressure and conjunctival hemorrhage occurred respectively in 2 eyes of the aflibercept group. There were no serious complications related to drug injection such as retinal detachment, complicated cataract, endophthalmitis and retinal pigment epithelial tear in the two groups, and there was no difference in the recurrence rate between the two groups(7% vs. 6%, P=1.000).CONCLUSION: On the basis of continuous 4 times of intravitreal injection, both conbercept and aflibercept are safe and effective in the treatment of wARMD, and the efficacy is even.

Effect and safety of aflibercept in the treatment of polypoidal choroidal vasculopathy with ranibizumab-resistant serous pigment epithelial detachment / 中华实验眼科杂志

Objective:To evaluate the effectiveness and safety of intravitreal injection of different doses of aflibercept for polypoidal choroidal vasculopathy (PCV) with serous pigment epithelial detachment (PED) resistant to ranibizumab.Methods:A non-randomized controlled clinical study was conducted.Seventy-three eyes of 73 patients with PCV and serous PED resistant to ranibizumab were enrolled at the First Affiliated Hospital of Zhengzhou University from January 2019 to December 2020.All patients were treated by intravitreal injection of 2 mg or 4 mg aflibercept according to patients' willingness.2 mg aflibercept or 4 mg aflibercept was intravitreally injected monthly for three consecutive months following pro re nata (PRN) regimen in 2 mg aflibercept group (38 eyes) and 4 mg aflibercept group (35 eyes), respectively.PED height and central macular thickness (CMT) were measured by optical coherence tomography, and the best corrected visual acuity (BCVA) was examined with a visual acuity chart and converted to logarithm of the minimum angle of resolution (LogMAR) unit before injection and 1 month, 2, 3, 6 months from the first injection.Intraocular pressure and treatment-related adverse events were recorded.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2021-KY-1252).Written informed consent was obtained from each patient prior to entering study cohort.Results:Thirty-three patients (86.84%) in 2 mg aflibercept group and 30 patients (85.71%) in 4 mg aflibercept group finished the treatment and follow-up, respectively. The PED, BCVA and CMT before treatment and at the end of follow-up were (379.24±95.50) and (280.09±120.50)μm, 0.68±0.27 and 0.51±0.19, (393.96±100.81) and (291.70±44.09)μm in 2 mg aflibercept group, respectively, showing statistically significant differences (all at P<0.05).The PED, BCVA and CMT before treatment and at the end of follow-up were (393.07±93.76) and (278.63±145.07)μm, 0.66±0.31 and 0.48±0.22, (377.43±79.61) and (284.67±84.88)μm in 4 mg aflibercept group, respectively, with statistically significant differences (all at P<0.05).The CMT value in 4 mg aflibercept group was significantly lower than that in the 2 mg aflibercept group in one month after injection ( P<0.05).No severe ocular and systemic adverse events were found during the follow-up, such as retinal detachment, endophthalmitis, cataract, and persistent high intraocular pressure. Conclusions:Both 2 mg and 4 mg aflibercept can effectively treat ranibizumab-resistant PCV with serous PED, and improve the anatomical structure of retina and BCVA.4 mg aflibercept can accelerate the recovery of PED and CMT.

Status and significance of BCR-ABL1 kinase domain mutations in imatinib-resistant chronic myelogenous leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia patients from Northeast China / 白血病·淋巴瘤

Objective:To explore the characteristics of BCR-ABL1 kinase domain mutations in imatinib-resistant chronic myelogenous leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph + ALL) patients from Northeast China and their impact on prognosis. Methods:The clinical data of 252 CML patients and 49 Ph + ALL patients who were admitted to the First Hospital of Jilin University from January 2013 to October 2018 were retrospectively analyzed. The samples of bone marrow or peripheral blood were collected from patients when imatinib treatment was not effective. Nested polymerase chain reaction (PCR) was used to amplify the BCR-ABL1 kinase domain, and Sequencing Analysis v5.4 software was used to analyze the mutation of BCR-ABL1 kinase domain. Patients were followed up for 6-48 months, and the survival analysis was performed. Results:Among 252 CML patients, the mutations in ABL1 kinase domain were found in 57 patients (22.6%), including 25 patients in the chronic phase, 21 patients in the accelerated phase and 11 patients in the blast crisis; 50 patients had 20 types of single point mutation, and the most common mutation types were E255K (16.0%, 8/50), T315I (14.0%, 7/50), M244V (8.0%, 4/50) and G250E (8.0%, 4/50), which were all concentrated in the P-loop and C-helix domains; 7 patients had double mutations; patients with multiple mutations had the worst prognosis, with a median overall survival (OS) time of 3.2 months. Among 49 Ph + ALL patients, 17 cases (34.7%) were positive for mutations in the BCR-ABL1 kinase domain, 14 patients had 12 types of single point mutation, and 3 patients had multiple mutations; the median OS time of patients with multiple mutations, mutations located in the P-loop and C-helix domains and mutations located in the other domains was 2.0, 8.0 and 18.0 months, and the difference in OS among the three groups was statistically significant ( P < 0.01). Conclusions:Among the imatinib-resistant CML and Ph + ALL patients from Northeast China, point mutations in the P-loop and C-helix domains are most commonly found. Multiple mutations, mutations in the P-loop and C-helix domains are related to the poor prognosis of the patients.

Observation of the curative effect of conbercept in the treatment of different types of diabetic macular edema / 中华眼底病杂志

Objective:To observe the efficacy of conbercept in the treatment of different types of diabetic macular edema (DME).Methods:A retrospective clinical study. From March 2019 to March 2021, 136 eyes of 136 patients with DME diagnosed in Department of Ophthalmology of Xi'an No.3 Hospital were included in the study. Among them, there were 65 males and 71 females; the average age was 56.65±8.65 years. All patients underwent best corrected visual acuity (BCVA), optical coherence tomography (OCT) examination, and glycosylated hemoglobin level (HbA1c) examination. Early Treatment Diabetic Retinopathy Study visual acuity chart was used for BCVA examination, which was converted into the logarithmic minimum angle of resolution (logMAR) visual acuity during statistics. An OCT instrument was used to measure the central retinal thickness (CRT) of the macula. According to the characteristics of OCT, DME was divided into diffuse retinal thickening (DRT) type, cystoid macular edema (CME) type, serous retinal detachment (SRD) type, mixed type, and grouped accordingly, respectively, about 30, 38, 33, 35 eyes. There was no significant difference in age ( F=1.189), sex ratio ( χ2=1.331), and HbA1c level ( F=3.164) of the four groups of patients ( P>0.05). All eyes were treated with intravitreal injection of 10 mg/ml conbercept 0.05 ml (including conbercept 0.5 mg) once a month for 3 consecutive times, and then treated as needed after evaluation. BCVA and OCT examinations were performed 1, 3, and 6 months after treatment with the same equipment and methods as before treatment. The changes of BCVA and CRT before and after treatment were compared and observed. For measurement data subject to normal distribution, one-way analysis of variance was performed for comparison between groups; χ2 test was performed for comparison of count data. Results:Before treatment, the logMAR BCVA of the eyes in the DRT group, CME group, SRD group, and mixed group were 0.68±0.11, 0.69±0.15, 0.71±0.12, 0.73±0.14, and CRT was 631.4±50.7, 640.6±55.7, 652.3±63.4, 660.4±61.8 μm. Compared with before treatment, 1, 3, 6 months after treatment, DRT group (BCVA: t=8.139, 11.552, 11.672; CRT: t=16.163, 21.653, 25.855), CME group (BCVA: t=8.923, 9.995, 13.842; CRT: t=16.163, 21.653, 25.855), SRD type group (BCVA: t=5.171, 7.315, 6.051; CRT: t=9.099, 13.731, 21.306), mixed type group (BCVA: t=5.072, 6.939, 7.142; CRT: t=6.920, 15.352, 17.538) The BCVA of the affected eyes was significantly increased, and the CRT was significantly decreased, and the difference was statistically significant ( P<0.05). At 6 months after treatment, the differences in logMAR BCVA and CRT of the 4 groups of eyes were statistically significant ( χ2=58.478, 64.228; P<0.05). The average number of injections in the eyes of the DRT group, CME group, SRD group, and mixed group were 3.37±1.35, 3.68±1.38, 4.18±1.40, 4.13±1.50 times, respectively. Compared with the average number of injections in the eye, the difference was statistically significant ( χ2=9.139, P=0.028). Conclusions:Conbercept can effectively reduce CRT and increase BCVA in eyes with different types of DME. Compared with SRD type and mixed type, DRT and CME type eye are more effective in improving vision, CRT reduction degree is greater, and the number of injections is less.

Association between single nucleotide polymorphism and response to anti vascular endothelial growth factor therapy in patients with wet age-related macular degeneration / 中华眼底病杂志

Objective:To observe the relationship between the response to anti-vascular endothelial growth factor (VEGF) drug treatment and single nucleotide polymorphism (SNP) genotype in patients with wet age-related macular degeneration (wAMD).Methods:A retrospective clinical study. From August 2019 to September 2020, 103 eyes of 103 wAMD patients diagnosed in Tianjin Medical University Eye Hospital were included in the study. Among them, there were 59 males (57.28%, 59/103) and 44 females (42.72%, 44/103); the average age was 68.74±7.74 years. The standard logarithmic visual acuity chart was used to detect the Best Corrected Visual Acuity of the affected eye and converted to the logarithmic minimum angle of resolution (logMAR) visual acuity during statistics. Optical coherence tomography was used to detect the central retinal thickness (CRT) of the affected eye. At the same time, the patient's high-density lipoprotein cholesterol (HDL-C) was tested. All eyes were treated with intravitreal injection of anti-VEGF drugs once a month for 3 months. Before the initial treatment, peripheral venous blood from the patient were collected. Interleukin-8 ( IL-8), complement C3 gene ( C3), complement factor H ( CFH), liver lipase ( LIPC), cholesterol ester transfer protein ( CETP), ATP binding cassette subfamily a member 1 ( ABCA1), lipoprotein lipase ( LPL), fatty acid desaturation gene cluster ( FADS1) SNP. According to gene frequency, genotypes are divided into wild type and mutant type were detected. Qualitative data such as the frequency difference of the genotype distribution in the clinical phenotype and the Hardy-Weinberg equilibrium of the genotype distribution were compared with the Chi-square test or Fisher's exact test. Results:There were fewer CRT responders in IL-8 rs4073 mutant (TA+AA) patients than wild-type (TT) [odds ratio ( OR)=0.310, 95% confidence interval ( CI) 0.106-0.910, P<0.05). Among them, after the drug stratification test, the proportion of patients with IL-8 rs4073 locus TT genotype in the conbercept treatment group was less CRT non-responders ( OR=0.179, 95% CI=0.034-0.960, P=0.033). Patients with LIPC rs2043085 mutant (CT+TT) with BCVA increased ≥0.2 logMAR are more likely than wild-type (CC) ( OR=3.031, 95% CI 1.036-8.867, P<0.05); HDL-C level was significantly lower Compared with wild type (CC), the difference was statistically significant ( t=2.448, P=0.016). There was no significant difference in logMAR BCVA and CRT between IL-8 rs4073, LIPC rs2043085 mutant and wild-type patients before treatment ( IL-8 rs4073: Z=-0.198, -1.651; P=0.843, 0.099; LIPC rs2043085: Z=-0.532, -0.152; P=0.595, 0.879). C3 rs 225066, CFH rs800292, CETP rs708272, ABCA1 rs1883025, FADS1 rs174547, LPL rs12678919 have no correlation with anti-VEGF drug treatment response. Conclusions:Patients with wAMD are treated with anti-VEGF drugs. Those with IL-8 rs4073 locus A genotype may be less responsive to CRT. LIPC rs2043085 locus T genotypes may be relatively more responsive to BCVA.

Short-term efficacy of primary treatment and dressing change with aflibercept for exudative age-related macular degeneration / 中华眼底病杂志

Objective:To observe the short-term effects of intravitreal injection of aflibercept (IVA) for initial treatment and dressing change on exudative age-related macular degeneration (eAMD).Methods:A retrospective clinical study. From June 2018 to February 2021, forty-nine eAMD eyes of 38 patients who underwent IVA treatment in Department of Ophthalmology of Central Theater Command Hospital of People’s Liberation Army were included in the study. Among them, there were 24 males with 29 eyes and 14 females with 20 eyes; the average age was 66.82±8.71 years. All affected eyes were treated with IVA. The initial loading dose was 2.0 mg, which was injected once a month for 3 consecutive months, followed by monthly review and treatment as needed. Of the 49 eyes, 26 eyes were initially treated (initial treatment group), they were diagnosed within 3 months of the first onset and followed by IVA treatment, and no intraocular drugs and surgery were performed from the onset to the first diagnosis. Twenty-three eyes were treated with drug exchange therapy (dressing change group), they received intravitreal injection of ranibizumab and/or conbercept more than 4 times 6 months before the replacement therapy, during which there was persistent interlaminar cystoid edema and/or subretinal fluid (SRF) in the macular area and no improvement in pigment epithelial detachment (PED). Before IVA treatment, there were no statistically significant differences in the best corrected visual acuity (BCVA), foveal thickness (CMT), PED height (PEDH), and PED volume (PEDV) of the two groups of eyes before IVA treatment ( P>0.05). The same equipment and methods as before treatment were used for related examinations, and the changes of BCVA, CMT, PEDH, PEDV and complications of the two groups of eyes were recorded in 1, 3, and 6 months after treatment. The comparison of BCVA, CMT, PEDH, and PEDV between the two groups were used repeated measures analysis of variance. Results:Six months after treatment, the number of IVA injections in the eyes of the initial treatment group and dressing change group were 4.15±0.73 and 4.39±0.72 times, respectively, and the difference was not statistically significant ( t=-1.141, P=0.260). The BCVA, CMT, PEDH, and PEDV of the the initial treatment group ( F=5.345, 22.995, 6.764, 5.425) and the dressing change group ( F=12.519, 15.576, 8.843, 9.406) were significantly improved compared before treatment with 1, 3, and 6 months. All were statistically significant ( P<0.05). There was no significant difference in BCVA, CMT, PEDH, and PEDV between the initial treatment group and the dressing group at each time point after treatment ( F=1.741, 0.069, 0.876, 3.455; P>0.05). During the follow-up period, none of the affected eyes had complications such as persistent intraocular pressure increase, endophthalmitis, and retinal pigment epithelial tear. Conclusions:IVA can improve eyesight of patients with eAMD and reduce CMT, PEDH, and PEDV. The initial treatment and dressing change have the same effect.

Overexpression of a fusion protein of 4-coumaroyl-CoA ligase and polyketide synthase for raspberry ketone production in Chlamydomonas reinhardtii / 生物工程学报

Raspberry ketones have important therapeutic properties such as anti-influenza and prevention of diabetes. In order to obtain raspberry ketone from Chlamydomonas reinhardtii, two enzymes catalyzing the last two steps of raspberry ketone synthesis, i.e. 4-coumaryl-CoA ligase (4CL) and polyketide synthase (PKS1), were fused using a glycine-serine-glycine (GSG) tripeptide linker to construct an expression vector pChla-4CL-PKS1. The fusion gene 4CL-PKS1 driven by a PSAD promoter was transformed into a wild-type (CC125) and a cell wall-deficient C. reinhardtii (CC425) by electroporation. The results showed the recombinant C. reinhardtii strain CC125 and CC425 with 4CL-PKS1 produced raspberry ketone at a level of 6.7 μg/g (fresh weight) and 5.9 μg/g (fresh weight), respectively, both were higher than that of the native raspberry ketone producing plants (2-4 μg/g).

Genetic engineering of crops for insect resistance: An overview

Phytophagous insect incidence is a serious threat for reduction of crop productivity globally. There is anestimation of one fourth of crop is being destroyed by insects annually. Indeed, the development of insectresistant crops is a great milestone in agriculture to increase crop yield and reduce pesticide dependency.Genetic engineering facilitates development of insect resistant crops by expressing bacterial d-endotoxins andvegetative insecticidal proteins and other plant genes like lectins, protease inhibitors, etc. In addition, RNAinterference and genome editing through CRISPR Cas9 also provides new solutions for the development ofinsect-resistant crops. The resultant genetically modified crops showed resistance against lepidopteran, dipteran, homopteran and coleopteran insects. The insect-resistant crops have made a significant economic impactworldwide in terms of higher yield and low pesticide usage. In this review, we focus on different strategies fordeveloping transgenics against insect pest control by expressing different insecticidal proteins in crops

Guia prático de atualização: medicamentos biológicos no tratamento da asma, doenças alérgicas e imunodeficiências / Practical update guide: biological drugs in the treatment of asthma, allergic diseases and immunodeficiencies

O presente guia apresenta revisão extensa sobre imunobiológicos utilizados, liberados e ainda sob estudo, para o tratamento da asma, doenças alérgicas e imunodeficiências. Além das características físico-químicas de alguns desses fármacos, são revisadas as indicações e os resultados de estudos clínicos realizados para avaliar eficácia e segurança. Separados por doença específica, são apresentados os principais agentes disponíveis e aprovados para utilização segundo as normas regulatórias nacionais.

This guide presents an extensive review of immunobiological drugs used, approved and/or under investigation for the treatment of asthma, allergic diseases and immunodeficiencies. In addition to the physicochemical characteristics of some of these drugs, their indications and results of clinical studies evaluating efficacy and safety are reviewed. The main agents available and approved for use in each specific disease according to national regulatory standards are presented.

Progress in molecular studies of Ph-positive and Ph-like acute lymphoblastic leukemia / 中国医师杂志

Acute lymphoblastic leukemia (ALL) is a highly heterogeneous disease. The prognosis of adolescents and adults with ALL is worse than that of children with ALL, mainly because of differences in biological characteristics. Molecular markers are of great value on risk stratification, prognosis evaluation, treatment guidance. The good news is with the deepening of studies on pathogenesis of ALL and the introduction of new technologies, novel molecular markers of prognostic relevance continue to be discovered. It was found that increased Sox4 expression was shown to correlate with accelerate leukemia progression. Unfortunately, resistance to medicine is associated with two cyclin-dependent kinase-inhibitor-2AB (CDKN2A/B) deletion.And the deletion of IKZF1 increase risk of relapse. Meanwhile, Ph-like acute lymphoblastic leukemia is characterized by a variety of genetic alterations that activate kinase or cytokine receptor signaling pathway. Identification of these molecular markers will provide important insights into the treatment strategies. This paper reviewed the advance in the Ph-positive ALL and Ph-like ALL.

Progress in molecular studies of Ph-positive and Ph-like acute lymphoblastic leukemia / 中国医师杂志

Acute lymphoblastic leukemia (ALL) is a highly heterogeneous disease.The prognosis of adolescents and adults with ALL is worse than that of children with ALL,mainly because of differences in biological characteristics.Molecular markers are of great value on risk stratification,prognosis evaluation,treatment guidance.The good news is with the deepening of studies on pathogenesis of ALL and the introduction of new technologies,novel molecular markers of prognostic relevance continue to be discovered.It was found that increased Sox4 expression was shown to correlate with accelerate leukemia progression.Unfortunately,resistance to medicine is associated with two cyclin-dependent kinase-inhibitor-2AB (CDKN2A/B) deletion.And the deletion of IKZF1 increase risk of relapse.Meanwhile,Ph-like acute lymphoblastic leukemia is characterized by a variety of genetic alterations that activate kinase or cytokine receptor signaling pathway.Identification of these molecular markers will provide important insights into the treatment strategies.This paper reviewed the advance in the Ph-positive ALL and Ph-like ALL.

Advances of bifunctional anti-PD-L1/TGF-β fusion protein M7824 / 国际肿瘤学杂志

With the development of programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) inhibitors,the second generation of combined immunosuppressive agents emerge as the times require.As a bifunctional anti-PD-L1/transforming growth factor-β (TGF-β) fusion protein,M7824 can antagonize PD-L1 pathway and trap TGF-β at the same time,which can effectively enhance the immune response and reduce the occurrence of immune escape and drug resistance.The drug has achieved remarkable results in many preclinical studies,however,the indications,safety and efficacy still need to be confirmed by large-scale clinical research data.

Infecciones del tracto urinario, inmunidad y vacunación / Urinary tract infections, immunity, and vaccination

Resumen Las infecciones del tracto urinario (ITU) se consideran como una de las principales causas de morbilidad en el mundo, y Escherichia coli uropatogénica (UPEC, por sus siglas en inglés) es el agente causal asociado a estas infecciones. La alta morbilidad generada por las ITU y la limitación de tratamientos debido al aumento de la resistencia bacteriana a los diversos antibióticos inducen la búsqueda de nuevas alternativas contra estas infecciones. El conocimiento que se ha generado acerca de la respuesta inmunitaria en el tracto urinario (TU) es importante para el desarrollo de estrategias efectivas en la prevención, el tratamiento y el control de las ITU. Los avances en las herramientas de biología molecular y bioinformática han permitido generar proteínas de fusión consideradas como biomoléculas potenciales para el desarrollo de una vacuna viable contra las ITU. Las adhesinas fimbriales (FimH, CsgA y PapG) de UPEC son factores de virulencia que contribuyen a la adherencia, la invasión y la formación de comunidades bacterianas intracelulares. Pocos estudios in vivo e in vitro han mostrado que las proteínas de fusión promueven una respuesta inmunitaria eficiente y de protección contra las ITU causadas por UPEC. Adicionalmente, la vía de inmunización intranasal con moléculas inmunogénicas ha generado una respuesta en la mucosa del TU en comparación contra otras vías de inmunización. El objetivo de esta revisión fue proponer un diseño de vacuna contra las ITU causadas por UPEC, describiendo el panorama general de la infección, el mecanismo de patogenicidad de la bacteria y la respuesta inmunitaria del huésped.

Abstract Urinary tract infections (UTI) are considered one of the main causes of morbidity worldwide, and uropathogenic Escherichia coli (UPEC) is the etiological agent associated with these infections. The high morbidity produced by the UTI and the limitation of antibiotic treatments promotes the search for new alternatives against these infections. The knowledge that has been generated regarding the immune response in the urinary tract is important for the development of effective strategies in the UTI prevention, treatment, and control. Molecular biology and bioinformatic tools have allowed the construction of fusion proteins as biomolecules for the development of a viable vaccine against UTI. The fimbrial adhesins (FimH, CsgA, and PapG) of UPEC are virulence factors that contribute to the adhesion, invasion, and formation of intracellular bacterial communities. The generation of recombinant proteins from fimbrial adhesins as a single molecule is obtained by fusion technology. A few in vivo and in vitro studies have shown that fusion proteins provide an efficient immune response and protection against UTI produced by UPEC. Intranasal immunization of immunogenic molecules has generated a response in the urinary tract mucosa compared with other routes of immunization. The objective of this review was to propose a vaccine designed against UTI caused by UPEC, describing the general scenario of the infection, the mechanism of pathogenicity of bacteria, and the immune response of the host.

Relationship between mechanism of SIRT1-mediated mitophagy in myocardium and Mfn2 in diabetic rats / 中华麻醉学杂志

Objective To evaluate the relationship between the mechanism of silent information regulator factor 2-related enzyme 1 (SIRT1)-mediated mitophagy in the myocardium and mitofusin 2 (Mfn2) in diabetic rats.Methods Twenty-four SPF healthy adult male Sprague-Dawley rats,weighing 210-220 g,were allocated into 3 groups (n =8 each) using a random number table:control group (group C),diabetes mellitus group (group DM) and diabetes mellitus plus SIRT1 activator SRT1720 group (group DM+SRT).Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.SRT1720 1 mg/kg was injected via the caudal vein for 7 consecutive days in group DM+SRT.The ultrasonic method was used to measure the parameters of heart function including left ventricular end-diastolic volume (LVEDV),left ventricular end-systolic volume (LVESV),left ventricular ejection fraction (LVEF),heart rate (HR),E wave velocity (E),A wave velocity (A) and E/A ratio.Myocardial specimens were obtained for determination of the interaction between SIRT1 and Mfn2 and acetylation of Mfn2 (by co-immuno-precipitation) and expression of SIRT1,Mfn2,microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ),LC3 Ⅰ,Beclin1 and P62 (by Western blot).The ratio LC3 Ⅱ to LC3 Ⅰ (LC3 Ⅱ / Ⅰ ratio) was calculated.Results Compared with group C,LVEDV,LVEF,HR,E and E/A ratio were significantly decreased,A was increased,LC3 Ⅱ / Ⅰ ratio was decreased,the expression of Beclin1,SIRT1 and Mfn2 was down-regulated,the expression of P62 was up-regulated,and the interaction between SIRT1 and Mfn2 was decreased in group DM (P＜0.05).Compared with group DM,LVEDV,LVEF,E and E/A ratio were significantly increased,A was decreased,LC3 Ⅱ / Ⅰ ratio was increased,the expression of Beclin1 and SIRT1 was up-regulated,the expression of P62 was down-regulated,the interaction between SIRT1 and Mfn2 was increased,and the acetylation of Mfn2 was decreased in group DM+SRT (P＜0.05).Conclusion The mechanism by which SIRT1 mediates mitophagy in the myocardium is related to SIRT1-induced deacetylation of Mfn2 in diabetic rats.

Recombinant human tumor necrosis factor receptor type Ⅱ : IgG Fc fusion protein combined with methotrexate for the treatment of psoriasis and their effects on levels of interleukin-17A and tumor necrosis factor-α / 中华皮肤科杂志

Objective To evaluate the effect of recombinant human tumor necrosis factor receptor type Ⅱ:IgG Fc fusion protein (rhTNFR:Fc,trade name Etanercept) combined with methotrexate on levels of interleukin-17A (IL-17A) and tumor necrosis factor-α (TNF-α) in the serum and mononuclear cells of patients with moderate to severe plaque psoriasis.Methods A total of 30 patients with moderate to severe plaque psoriasis were enrolled from Department of Dermatology of Tenth People's Hospital of Tongji University between August 2014 and February 2016,and then were randomly and equally divided into Etanercept group and Etanercept + methotrexate group.The treatment lasted 24 weeks.Fifteen healthy blood donors served as healthy control group.Enzyme-linked immunosorbent assay (ELISA) and real-time quantitative PCR were performed to measure the serum levels and mRNA expression of IL-17A and TNF-α,respectively,in the patients of the above two groups before and after the treatment.Results Before the treatment,the serum levels of IL-17A and TNF-ct,as well as the mRNA expression of IL-17A and TNF-α in peripheral blood mononuclear cells (PBMCs),were all significantly higher in all the patients than in the healthy controls (all P ＜ 0.05).After the treatment,compared with the Etanercept group,the Etanercept + methotrexate group showed significantly lower serum levels of IL-17A (142.67 ± 14.82 vs.163.54 ± 23.18,P ＜ 0.05) and TNF-α (70.07 ± 25.02 vs.91.98 ± 14.62,P ＜ 0.05),as well as lower mRNA expression of IL-17A (1.12 ± 0.33 vs.1.56 ± 0.77,P ＜ 0.05) and TNF-α in PBMCs (2.50 ± 1.04 vs.3.61 ± 2.14,P ＜ 0.05).Conclusion Etanercept combined with methotrexate is superior to Etanercept alone in the treatment of psoriasis,and can reduce treatment duration and improve therapeutic effect,likely by down-regulating the expression of IL-17A and TNF-α.

Role of PKCα/HO-1 signaling pathway in endotoxin-induced damage to alveolar macrophages of rats: the relationship with mitofusin-1 / 中华麻醉学杂志

Objective To evaluate the role of protein kinase Cα (PKCα)/heme oxygenase-1 (HO-1) signaling pathway in endotoxin-induced damage to alveolar macrophages and the relationship with mitofusin-1 (Mfn1) in rats.Methods Rat alveolar macrophages NR8383 cells cultured in vitro were seeded in 96-well plates at a density of 1 × 104 cells/ml.NR8383 cells were divided into 5 groups (n =15 each)using a random number table:control group (group C),endotoxin challenge model group (group E),PKCα inhibitor Go6976 group (group G),PKCα agonist PMA group (group P) and dimethyl sulfoxide group (group D).NR8383 cells were stimulated with 10 μg/ml lipopolysaccharide (LPS) to establish the model of endotoxin challenge in alveolar macrophages.In G,P and D groups,cells were pretreated with 5 μmol/L Go6976,100 nmol/L PMA and 0.1％ dimethyl sulfoxide,respectively,for 30 min starting from 30 min before stimulation with LPS,and 10 μg/ml LPS was then given.The cells were collected after 24 h of incubation for measurement of malondialdehyde (MDA) and reactive oxygen species (ROS) contents,superoxide dismutase (SOD) activity and expression of PKCα,HO-1 and Mfn1 protein and mRNA (by fluorescent quantitative polymerase chain reaction or Western blot).Results Compared with group C,MDA and ROS contents were significantly increased,the SOD activity was decreased,the expression of PKCα and HO-1 protein and mRNA was up-regulated,and the expression of Mfn1 protein and mRNA was downregulated in E,G,P and D groups (P＜0.05).Compared with group E,MDA and ROS contents were significantly increased,the SOD activity was decreased,and the expression of PKCα,HO-1 and Mfn1 protein and mRNA was down-regulated in group G,MDA and ROS contents were significantly decreased,the SOD activity was increased,and the expression of PKCα,HO-1 and Mfn1 protein and mRNA was upregulated in group P (P＜0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Conclusion Promotion of Mfn1 expression following PKCα/HO-1 signaling pathway activation is the endogenous protective mechanism of endotoxin-induced damage to alveolar macrophages of rats.

Effects of RNAi targeting CDX2 gene expression on biological characters of human leukemia cells and its relevant mechanisms / 天津医药

Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase-9, Bax mRNA and protein were tested by RT-PCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase-9 and Bax mRNA and protein were significantly higher than those of other two groups (P<0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the down-regulation of BCR-ABL and the up-regulation of caspase-9 and Bax.

Effect of AGEs on expression of mitochondrial fusion proteins Mfn1 and Mfn2 in cultured human aortic endothelial cells / 中国病理生理杂志

AIM: To clarify whether advanced glycation end products (AGEs) can influence the expression of mitochondrial fusion proteins Mfn1 and Mfn2 in cultured human aortic endothelial cells (HAECs) in vitro.METHODS:AGE-BSA was used as AGEs.Purchased primary human aortic endothelial cell line was multiplied, and transferred to dif-ferent passages for subsequent grouping.For dose-dependent experiment, HAECs were divided into 4 groups, and the con-centrations of AGE-BSA in each group were 0 mg/L (control group), 50 mg/L, 100 mg/L and 200 mg/L, respectively. For time-dependent experiment, HAECs were divided into 5 groups with the same concentration (100 mg/L) of AGE-BSA, but the intervention time was 0 h (control group), 6 h, 12 h, 24 h and 48 h, respectively.The mRNA and protein expres-sion levels of Mfn1 and Mfn2 in the HAECs were detected by real-time PCR and Western blot, respectively.RESULTS:Exposure of the HAECs to AGEs at different concentrations for 24 h all down-regulated the mRNA and protein expression levels of Mfn1 and Mfn2.Except for 6 h intervention group, 100 mg/L AGEs intervention for 12 h, 24 h and 48 h all down-regulated the mRNA and protein expression levels of Mfn1 and Mfn2 in cultured HAECs.CONCLUSION: AGEs down-regulates the expression of mitochondrial fusion proteins Mfn1 and Mfn2 in cultured HAECs, indicating that AGEs may influence mitochondrial dynamics of human aortic endothelial cells.

Effect of heme oxygenase-1 protein transduction on acute lung injury in septic rats / 中华麻醉学杂志

Objective To evaluate the effect of heme oxygenase-1 (HO-1) protein transduction on acute lung injury in septic rats.Methods Eighteen healthy male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly allocated into 3 groups (n =6 each) using a random number table:sham operation group (group Sham),sepsis group (group Sep),and fusion protein PEP-1-HO-1 group (group HO).Sepsis was produced by cecal ligation and puncture (CLP).In group HO,PEP-1-HO-1 fusion protein 0.6 mg was injected via the left iliac vein at 1 h before CLP and 5 h after CLP.The equal volume of normal saline was given instead of PEP-1-HO-1 in Sham and Sep groups.At 12 h after CLP,blood samples were collected from the right common carotid artery for measurement of serum tumor necrosis factoralpha (TNF-α) and interleukin-6 (IL-6) concentrations.The rats were then sacrificed,and lungs were removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio) and malondialdehyde (MDA) content (by thiobarbituric acid colorimetric method).Results Compared with group Sham,the W/D ratio and MDA content were significantly increased,the serum TNF-α and IL-6 concentrations were significantly increased (P＜0.05),and the pathological changes were significantly aggravated in Sep and HO groups.Compared with group Sep,the W/D ratio and MDA content were significantly decreased,the serum TNF-α and IL-6 concentrations were significantly decreased (P＜0.05),and the pathological changes were significantly attenuated in group HO.Conclusion HO-1 protein transduction can attenuate acute lung injury in septic rats,and the mechanism is probably related to inhibition of lipid peroxidation in lung tissues and systemic inflammatory responses.

Prokaryotic expression and purification of antimicrobial peptide LL-37 and the inhibiting effect against Candida albicans / 中华妇产科杂志

Objective To study the inhibitory effect of antimicrobial peptide LL-37 on Candida albicans through its ability to promote the secretion of immune factors by vaginal epithelial cells. Methods (1) LL-37 prokaryotic expression vector pET-Duet/LL-37 was constructed and its expression was induced in Escherichia coli M15. The expressed LL-37 fusion protein was purified and identified by western blot. Antifungal activity of the purified protein was initially identified by Kirby-Bauer (K-B) method. (2) Purified LL-37 protein was added to human vaginal epithelial cells co-cultured with Candida, and inhibitory effect on Candida growth was determined by the glucose consumption method. Interferon γ (IFN-γ), interleukin 10 (IL-10) concentration and IFN-γ/IL-10 ratio were measured by ELISA at different time points. Results (1) LL-37 fusion protein was purified to 96% purity at a concentration of 433.92 μg/ml, and was shown to possess anti-fungal activity confirmed by the K-B method. (2) A Candida-vaginal epithelial cells co-culture system was successfully constructed. LL-37 recombinant protein inhibited the growth of Candida with absorbance values significantly higher in the treatment group compared to the control group at all measured time points (12-hour:3.008±0.003 versus 2.967±0.003, 24-hour:2.941±0.003 versus 2.601±0.003, 48-hour:2.893 ± 0.004 versus 2.409 ± 0.003; all P<0.01). Furthermore, the rate of decrease was also much slower compared to the control group. In both control and experimental groups, IFN-γ and IL-10 secretion levels were observed to rise at first peaking at 24 hours and subsequently decrease. For each time period, IFN-γconcentration in the experimental group was significantly higher at 24 hours compared to the control group [(104.00 ± 1.07) versus (85.17 ± 0.28) pg/ml, P<0.01]. In contrast, IL-10 concentrations were significantly lower than the control group at all time points (P<0.01). IFN-γ/IL-10 ratio was also observed to be significantly higher than the control group at all measured time points (P<0.01). Conclusions (1) Recombinant protein LL-37 could significantly inhibit the growth of Candida. (2) By influencing the secretion of immune factors such as IFN-γ, IL-10, etc, recombinant protein LL-37 is able to adjust vaginal epithelial cells local immunity, and enhance resistance to Candida infection.