Software educativo para la asignatura Estratificación de Riesgos Medioambientales / Educational software for the subject Stratification of Environmental Risks

Se realizó una investigación relacionada con la innovación tecnológica, en la Facultad de Enfermería-Tecnología de la Salud de Santiago de Cuba, durante el curso escolar 2019-2020, con el objetivo de diseñar un software educativo para la asignatura Estratificación de Riesgos Medioambientales, dirigido a los estudiantes de técnico medio en Vigilancia y Lucha Antivectorial. Se utilizaron los métodos teóricos: análisis-síntesis, histórico-lógico, modelación, sistémico-estructural e inductivo-deductivo; y empíricos: observación y análisis documental. La muestra fue de 44 estudiantes y 6 profesores escogidos al azar. Se concluye que el software propuesto es factible, pertinente y necesario como medio de enseñanza; proporciona información actualizada, su navegación es fácil y amena, y permite la autoevaluación de los estudiantes al interactuar con él mismo, lo que contribuye a mejorar el trabajo independiente.

An investigation related to the technological innovation was carried out in the Health Nursing-Technology Faculty from Santiago de Cuba, during the school course 2019-2020, aimed at designing an educational software for the subject Stratification of Environmental Risks, directed to medium technician students in Surveillance and Vector Control. The theoretical methods used were: analysis-synthesis, historical-logical, modelation, systemic-structural and inductive-deductive; and the empiric methods were: observation and documental analysis. The sample had 44 students and 6 professors chosen at random. It was concluded that the proposed software is feasible, pertinent and necessary as teaching aid; provides up-to-date information, it is easy and interesting to surf internet, and allows the self-appraisal of students in the interaction with themselves, what contributes to improve the independent work.

Estudio observacional de las características clínicas, epidemiológicas y de laboratorio en pacientes pediátricos con dengue de la ciudad de Córdoba / Observational study of clinical, epidemiological, and laboratory characteristics of pediatric patients with dengue in the city of Córdoba

Introducción. El dengue es la enfermedad transmitida por mosquitos con mayor propagación mundial en los últimos años. Presenta un amplio espectro de manifestaciones clínicas y, en ocasiones, evoluciona a un estado crítico llamado dengue grave. Su tratamiento es de sostén. La información disponible acerca de las características clínicas, epidemiológicas y de laboratorio de la enfermedad en la población pediátrica es limitada. Objetivo. Describir la epidemiología y las manifestaciones clínicas y de laboratorio de la enfermedad. Población y métodos. Estudio descriptivo, observacional y retrospectivo. Incluyó pacientes entre 1 y 180 meses asistidos por dengue probable o confirmado en un hospital de niños, desde el 01 de enero de 2020 hasta el 31 de mayo de 2020. Resultados. Se incluyeron 85 pacientes por criterios microbiológicos de positividad o clínicoepidemiológicos. Veinticinco (29 %) confirmados por RT-PCR, todos serotipos DENV-1. La mediana de  edad fue de 108 meses (rango intercuartílico: 84-144). Las principales manifestaciones clínicas fueron fiebre, cefalea y mialgias. Los hallazgos de laboratorio más importantes fueron leucopenia, trombocitopenia y elevación de transaminasas. Conclusión. El reconocimiento y la comprensión de las alteraciones clínicas y de laboratorio que se presentan durante la enfermedad pueden permitir un abordaje eficaz y contribuir a la reducción de cuadros clínicos más graves en los niños.

Introduction. Dengue has been the most widespread mosquito-borne disease worldwide in recent years. It develops with a broad spectrum of clinical manifestations and sometimes progresses to a critical condition known as severe dengue. It is managed with supportive treatment. Available information about its clinical, epidemiological, and laboratory characteristics in the pediatric population is limited. Objective. To describe the clinical, epidemiological, and laboratory characteristics of dengue. Population and methods. Descriptive, observational, and retrospective study. It included patients aged 1 to 180 months seen due to probable or confirmed dengue at a children's hospital between 1/1/2020 and 5/31/2020. Results. A total of 85 patients with positive microbiological or clinical-epidemiological criteria were included. Of these, 25 (29%) were confirmed by RT-PCR; all corresponded to DENV-1 serotype. Patients' median age was 108 months (interquartile range: 84­144). The main clinical manifestations were fever, headache, and myalgia. The most important laboratory findings were leukopenia, thrombocytopenia, and high transaminase levels. Conclusion. The recognition and understanding of clinical and laboratory alterations that occur during dengue disease may allow an effective approach and help to reduce the more severe clinical form in children.

Caso importado de malaria por Plasmodium vivax en El Salvador. Un abordaje epidemiológico / Imported Case of Malaria by Plasmodium vivax in El Salvador. An Epidemiological Approach

Presentación del caso. Paciente masculino de origen guatemalteco con historia de fiebre alta de tipo intermitente, mialgias, artralgias, debilidad generalizada, mareo y vómito de contenido gástrico. Fue tratado inicialmente en un hospital privado con diagnóstico de síndrome febril agudo y referido a un hospital de la red nacional con diagnóstico de dengue con signos de alarma, al tercer día de estancia hospitalaria se diagnostica como un caso de malaria importado por Plasmodium vivax. Intervención terapéutica. Se le dio tratamiento antimalárico con cloroquina y primaquina. Evolución clínica. Presentó mejoría clínica y las pruebas de laboratorio de control reportaron resultados negativos para Plasmodium vivax

Case presentation. Male patient of Guatemalan origin with history of intermittent high fever, myalgia, arthralgia, generalized weakness, dizziness, and vomiting of gastric contents. He was initially treated in a private hospital with a diagnosis of acute febrile illness and referred to a national network hospital with a diagnosis of dengue with warning signs. On the third day of hospital stay a diagnosis of an imported malaria case by Plasmodium vivax was presented. Treatment. The patient was given antimalarial treatment consisting of chloroquine and primaquine. Outcome. The patient presented clinical improvement, and control laboratory tests were negative for Plasmodium vivax.

Efficacy, effectiveness, and safety of COVID-19 vaccine compared to placebo in preventing COVID-19 infection among 12-17 years old: A systematic review

Objectives@#The World Health Organization recently revised their recommendations and considered healthy children and adolescents as low priority group for COVID-19 vaccine. This review comprehensively assessed existing clinical evidence on COVID-19 vaccine in 12-17 years old.@*Methods@#Included in this review were any type of study that investigated the efficacy, immunogenicity, safety, and effectiveness of COVID-19 vaccine on protection against SARS-COV-2 infection in 12-17 years old. Various electronic databases were searched up to March 15, 2023. Studies were screened, data extracted, risk of bias appraised, and certainty of evidence was judged using GRADE. Review Manager 5.4 was used to estimate pooled effects. Difference between the two groups was described as mean difference for continuous variables and as relative risk or odds ratio for categorical variables.@*Results@#There were six randomized controlled trials and 16 effectiveness studies (8 cohorts and 8 case control). Low certainty evidence showed that BNT162b2 (Pfizer) was effective, immunogenic, and safe in healthy adolescents. There were 15 effectiveness studies on BNT162b2 (Pfizer) in healthy adolescent and one on immunocompromised patients. It was protective against infection with any of the variants, with higher protection against Delta than Omicron. BNT162b2 is protective against hospitalization and emergency and urgent care (high certainty); and critical care and MIS-C (low). Very low certainty evidence noted that BNT 162b2 was also immunogenic in 12-21 years old with rheumatic diseases while on immunomodulatory treatment but with possible increased exacerbation of illness. Low certainty evidence demonstrated that mRNA-1273 (Moderna) was effective, immunogenic, and safe. Low to very low certainty evidence were noted on the safety and immunogenicity of two vector base vaccines (ChAdOx1-19 and Ad5 vector COVID vaccine) and two inactivated vaccines (CoronaVac and BBIBP CorV).@*Conclusion@#There is presently low certainty evidence on the use of RNA vaccines in 12-17 years old. The recommendation on its use is weak. There is presently insufficient evidence for the use of inactivated and vector-based COVID-19 vaccines. Different countries should consider whether to vaccinate healthy adolescent without comprising the other recommended immunization and health priorities that are crucial for this age group. Other factors including cost-effectiveness of vaccination and disease burden should be accounted.

Significance and successful construction of stable transfection of MH7A cells with lower TRAF2 expression using lentiviral vector / 安徽医科大学学报

Objective To construct a stable synovial cell line MH7A from rheumatoid arthritis(RA)patients using lentiviral vectors that interfere with the expression of tumor necrosis factor receptor associated factor 2(TRAF2),and to study the role of TNF-α-TRAF2 signaling in MH7A abnormal proliferation.Methods Based on the design principles of human TRAF2 gene sequence and shRNA sequence,three pairs of TRAF2 shRNA interference se-quences were designed and synthesized.The primers were annealed by PCR,and a linear vector was obtained by double enzyme digestion PLKO.1-puro.The linearized vector was connected to the annealed primers through Solu-tion I,and the connected products were introduced into receptive cells.The plates were coated,and positive colo-nies were selected for sequencing.Three different recombinant plasmids of PLKO.1-TRAF2-shRNA lentivirus were constructed,and lentivirus packaging plasmids was used to package logarithmic growth phase HEK 293T cells.Vi-rus solution was collected to infect MH7A cells.At the same time,puromycin was used to screen MH7A stable transgenic strains with low TRAF2 expression.CCK-8 method,Western blot,and qPCR were used to detect the proliferation function of MH7A induced by TNF-α and low expression of TRAF2,as well as downstream signal TRAF2,P65 protein expression and mRNA levels.Results PLKO.1-TRAF2-shRNA(1),PLKO.1-TRAF2-shR-NA(2),and PLKO.1-TRAF2-shRNA(3)lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were successfully constructed.The three TRAF2-shRNA lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were respectively introduced into the lentivirus packaging plasmid of HEK 293T to obtain virus solution.After infecting MH7A cells with the virus solution,they were treated with puromycin(2.00 μ G/mL)screening and obtaining MH7A stable transgenic plants after 2 days.Through qPCR and Western blot results,it was found that the expression of TRAF2 mRNA and protein in PLKO.1-TRAF2-shRNA(1)MH7A stably transfected cells was significantly reduced compared to the negative control group.The results of CCK-8 and Western blot showed that after knocking down TRAF2 in MH7A,the proliferation of MH7A cells with low TRAF2 expression induced by TNF-α and the phosphorylation level of P65 were significantly reduced.Conclusion A sta-ble transgenic strain of PLKO.1-TRAF2-shRNA(1)MH7A cells was successfully constructed to investigate the role of TNF-α-TRAF2 signal activation in mediating abnormal proliferation of RA synovial cells.

Construction and drug resistance evaluation of cell strains which stably express FIP1L1-PDGFRA protein and its mutants / 基础医学与临床

Objective To construct mouse BaF3-FIP1L1-PDGFRA(F/P),BaF3-F/P-T674I and BAF3-F/P-D842V pre-B cell strains which stably express F/P fusion protein and its T674I and D842V mutants in order to e-valuate their activity by checking their responses to tyrosine kinase inhibitors(TKIs).Methods Lentivirus infected technique was used to transfect the target gene into BaF3 cells.RT-qPCR was used to detect mRNA expression,and CCK-8 method was used to detect the inhibitory effect of TKIs on the proliferation of stable cell strains.Results The constructed BaF3-F/P,BaF3-F/P-T674I and BAF3-F/P-D842V cell strains all transcripted FIP1L1 and PDGFRA mRNA.They exhibited malignant phenotypic characteristics of proliferation independent of IL-3 and sen-sitivity to corresponding TKIs.Conclusions The pre-B-cell strains stably expressing F/P and its T674I and D842V mutants are successfully constructed,which provide a good cell model for the development of compounds targeting at those molecules.

Research and Application of Drug Pair Extraction Method Integrating Co-occurrence and Semantic Information / 世界科学技术-中医药现代化

Objective To propose a drug pair extraction algorithm integrating co-occurrence and semantic information for prescription data.Methods The prescription data were transformed into matrix data,and the association information between drugs was calculated as the initial screening index.Then the word vector was constructed based on the prescription data,and the semantic similarity between drugs was calculated as the second screening index,so as to extract potential drug pairs.The algorithm of this paper and the classical Apriori algorithm were experimented on 1090 lung cancer outpatient prescriptions respectively,and the experimental extraction results were compared and analyzed,so as to verify the usability and effectiveness of this drug pair extraction algorithm.Results Compared with the Apriori algorithm,the present algorithm had better effect in extracting drug pairs,which could reasonably help to narrow down the range of options of potential drug pairs under the situation of large difference in drug frequencies,and successfully extracted 88 groups of drug pairs in medical cases under the range of recommended threshold settings.Conclusion The method of word frequency combined with semantic information for extracting potential drug pairs is feasible and effective,and can provide methodological reference for experience mining in clinical prescription medication.

Construction and clinical practice of vector management strategy for preventing ventilator-associated pneumonia by blocking etiological characteristics / 中国实用护理杂志

Objective:To establish an environmental management strategy for the prevention of ventilator-associated pneumonia from the perspective of etiological characteristics and to verify its application effect.Methods:Based on a sampling survey, this study constructed preventive management strategies for ventilator-associated pneumonia by blocking pathogen characteristics from the perspective of both colonization and infection management in patients. From July 2021 to June 2023, a non-synchronous randomized controlled study was conducted, including a control group of 59 cases and an experimental group of 57 cases from ICU of Tianjin Teda Hospital, all of them were mechanically ventilated patients. The effectiveness of the strategy was confirmed.Results:In the control group, there were 35 males and 24 females, with an average age of (46.97 ± 18.84) years. In the experimental group, there were 39 males and 18 females, with an average age of (47.49 ± 13.85) years. During the study period, there were 9 cases of ventilator-associated pneumonia (VAP) in the control group and 2 cases in the experimental group, the difference between the two groups was statistically significant (exact odds ratio=0.031). The duration of mechanical ventilation in the experimental group (122.41 ± 18.36) h, which was shorter than that in the control group (187.62 ± 18.05) h, and the difference was statistically significant ( t=19.28, P<0.05). The length of ICU stay in the experimental group was (8.38 ± 0.79) d, in the control group was (10.99 ± 1.10) d, the difference between them was statistically significant ( t=14.66, P<0.05). On the 7th day, there were 7 cases of positive pathogenic bacteria in sputum culture in the experimental group, which was significantly different from the 29 cases in the control group ( χ2=16.73, P<0.05). Conclusions:The vector management strategy for preventing ventilator-associated pneumonia by blocking etiological characteristics can reduce the incidence of VAP, shorten the duration of mechanical ventilation and ICU stay, and reduce the pathogen load in the sputum of mechanically ventilated patients on the 7th day.

Experimental study on treatment of severe limb ischemia with Ad-hVEGF-hHGF gene / 实用医学杂志

Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue.Methods 84 Kunming mice were randomly divided into sham group,control group,VEGF group,HGF group and VEGF+HGF group,and the left lower limb ischemia model was established.The blood supply of ischemic tissue was observed by rheometer,and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA.Immunohistochemical staining was used to detect angiogenesis(CD31,SMA)in ischemic tissues.Safety was assessed by side effects during treatment in mice.Results After the successful modeling,the blood flow velocity of the left lower limb in each group decreased significantly.On the 7th day after operation,the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation(P<0.05),and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups(P<0.05).On the 28th day after operation,the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized,the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups,and both VEGF group and HGF group were significantly better than the control group(P<0.05).On the 7th,14th,and 28th days following surgery,HGF and VEGF protein levels in the Ad-HGF,Ad-VEGF,and Ad-VEGF-HGF groups were substantially greater than those in the control group(P<0.05).The expression level in the Ad-VEGF-HGF group peaked on the 14th day(all P<0.001)and subsequently declined to preoperative levels on the 28th day after operation.Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level,encour-aging angiogenesis after lower limb ischemia,increasing blood flow,and improving lower limb circulation.

Removal of the selective marker gene in the fusion gene expression vector of plant anti-caries vaccine / 中国组织工程研究

BACKGROUND:In consideration of the food safety and ecological safety of transgenic plants,the retention of marker genes is the primary safety issue affecting transgenic plants. OBJECTIVE:Based on the principle of immune caries prevention,our research team successfully constructed the plant anti-caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 for these two caries causing virulence factors surface protein and glucosyltransferase,which provides a basis for the research and development of transgenic plant vaccine. METHODS:In this study,the selective marker genes Km and GUS in the plant caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 were removed by DNA recombination technology through a series of steps such as DNA fragment separation,connection,transformation,clone detection,and sequencing. RESULTS AND CONCLUSION:The efficiency of marker gene removal was 99%.This study has laid a good experimental foundation for the safe production of transgenic plant vaccine against dental caries,and also provided ideas for the construction of other plant vaccine vectors.

Differentiation of insulin-producing cells from human umbilical cord mesenchymal stem cells infected by MAFA-PDX1 overexpressed lentivirus / 中国组织工程研究

BACKGROUND:Transplantation of stem cell-derived islet β cells has been considered effective for the treatment of type 1 diabetes.Human umbilical cord mesenchymal stem cell is an ideal cellular source,but with a low differentiation efficiency to islet β cells. OBJECTIVE:To explore the possibility of human umbilical cord mesenchymal stem cells modified by MAFA and PDX1 to differentiate into insulin-producing cells. METHODS:MAFA-PDX1 lentivirus expression vectors were constructed.The efficiency and potentiality of human umbilical cord mesenchymal stem cells differentiated into insulin-producing cells with three methods were compared by cell morphology,RT-qPCR,and dithizone staining[protocol A:Simple lentivirus group;protocol B:Drug(nicotinamide β-mercaptoethanol)induction followed by lentivirus group;protocol C:lentivirus and drug induction group]. RESULTS AND CONCLUSION:(1)Morphological change of cells:Cell morphology was all altered after the induction of three protocols.At day 11,human umbilical cord mesenchymal stem cells induced by protocol B showed the most cell clusters among the three protocols,appearing aggregated islet-like cell clusters.(2)Islet-related gene expression detected by RT-qPCR:Horizontal comparison of the three protocols at the same induction time point showed that the expression levels of MAFA and PDX1 genes were the highest in protocol C on day 5 of induction,and those in protocol B were the highest on day 11 of induction.Human umbilical cord mesenchymal stem cells induced by protocol B had the greatest expression of GCG gene at day 5,INS and GLUT2 genes at day 11.(3)Dithizone staining to identify zinc ions:parts of the post-induced cells were stained brownish red by dithizone on day 11.The partial small island cells were stained brownish red with a darker color(positive expression)in protocol B.(4)It is concluded that the overexpression of MAFA and PDX1 can promote the differentiation of human umbilical cord mesenchymal stem cells into insulin-producing cells.The combination of MAFA-PDX1 gene modification and drug induction is superior to the single gene modification.

Construction of a BAC Vector and Its Application for Haplotype Analysis of CYP2A6 / 医学研究杂志

Objective To construct one high-copy BAC vector with cloning capacity for large DNA fragment insertion and use it for the haplotype analysis of CYP2A6 gene.Methods Oligos including multiple cloning sites were annealed and ligated into the Hind Ⅲ/BamH Ⅰ site of pGEM-3Z and pBeloBACll,separately.Then,the intermediate vectors were digested by Hind Ⅲ and ligated together,so as to get the head-to-tail oriented high-copy BAC vector pBAC-BJH after the blue-white spotting test.STI PCR method was used for the amplification of whole CYP2A6 gene,and purified amplicon was double digested by BstB Ⅰ/Mlu Ⅰ and ligated into the same site of newly constructed vector pBAC-BJH.Several clones were then picked up and sequenced for the haplotype analysis of carriers with newly discovered CYP2A6 variant 355A＞T.Results One high-copy BAC vector pBAC-BJH with 7 newly added multiple cloning sites was successfully constructed.High copy number and multiple cloning sites were the advantages of this plasmid.With this vector,haplotype analysis result for 22C＞T,51A＞G,355A＞T in carrier with newly detected CYP2A6mutation was CGT.Conclusion One vector pBAC-BJH convenient for cloning large DNA fragment was successfully developed,and it can be used for the haplotype analysis of cytochrome p450 gene and cloning or sequencing of other genes with large genomic DNA inserts.

Construction of recombinant adenovirus vector of Uhrf1 gene and its role in DNA damage repair of cardiomyocytes / 中国病理生理杂志

AIM:To construct a recombinant adenovirus vector carrying the mouse ubiquitin-like with plant homeodomain and RING finger domains 1(Uhrf1)gene,validate the expression of Uhrf1 in neonatal mouse cardiomyo-cytes and explored its role in hydrogen peroxide(H2O2)-induced DNA damage.METHODS:The mouse Uhrf1 gene cod-ing sequence was amplified by polymerase chain reaction(PCR),digested,and inserted into the pADM-CMV-C-FH vec-tor to create the recombinant adenoviral plasmid ADM-Uhrf1.Following transfection into HEK293T cells,we generated re-combinant adenoviral particles,amplified,purified,and determined the titer.Neonatal mouse cardiomyocytes were infect-ed at an multiplicity of infection(MOI)of 50,UHRF1 protein expression was validated via Western blot and immunofluo-rescence staining.H2O2-induced DNA damage was explored along with adenovirus-mediated Uhrf1 overexpression to inves-tigate its role in DNA damage repair.RESULTS:ADM-Uhrf1 virus titer,determined by capsid immunofluorescence as-say,was 1.8×1013 pfu/L.Western blot confirmed a significant increase in UHRF1 protein expression(P＜0.05),with im-munofluorescence indicating predominant nuclear localization.Uhrf1 overexpression effectively inhibited the expression of the DNA damage marker,phosphorylated H2AX protein(γH2AX)(P＜0.01).CONCLUSION:We successfully con-structed a recombinant adenoviral vector carrying the mouse Uhrf1 gene,facilitating Uhrf1 overexpression in neonatal mouse cardiomyocytes.Furthermore,this overexpression effectively alleviated DNA damage in cardiomyocytes.

Intranasal immunization with single-dose vaccine based on recombinant influenza virus H1N1 expressing the extracellular domain of respiratory syncytial virus G protein induces robust immunity and protection in mice / 中华微生物学和免疫学杂志

Objective:To construct a novel respiratory syncytial virus (RSV) vaccine based on a recombinant influenza virus vector and evaluate its immune protective effects in mice.Methods:A recombinant H1N1 influenza A virus (IAV) expressing the extracellular domain (Gecto) of RSV A2 G protein was constructed and rescued, named as PR8NAGecto/WSN. After in vitro verification of the Gecto expression and PR8NAGecto/WSN growth kinetics, a single dose of PR8NAGecto/WSN was used to immunize BALB/c mice through intranasal administration to evaluate the efficacy of PR8NAGecto/WSN by assessing humoral (IgG, neutralizing antibody), mucosal (IgA) and cellular immunity (IFN-γ ELISPOT). Four weeks after immunization, the mice were challenged with RSV A2 or RSV B9320 to evaluate the protective effects of PR8NAGecto/WSN by analyzing mouse body weight changes, lung tissue virus titers and pathological changes. Results:A single-dose intranasal immunization with PR8NAGecto/WSN induced robust humoral, mucosal and cellular immunity in mice. Moreover, the mice in the immunized group had lower lung virus loads and mild lung pathological damages following the challenge with RSV A or RSV B subtype as compared with the control group.Conclusions:A single-dose intranasal immunization with PR8NAGecto/WSN induces robust immunity and provide protection against RSV A and B challenges in mice. This study provides new ideas and reference for the development of novel mucosal vaccines against RSV.

False-positive HIV-1 nucleic acid testing results in patients with severe thalassemia after receiving cell and gene therapy / 中华检验医学杂志

A 11-year old female patient with severe thalassemia, receipt a lentivirus-based cell and gene therapy (CGT) therapy in Shenzhen Children′s Hosptial on July 27th, 2021. At the two follow-up visits after discharge, patient were continuously tested positive for HIV screening through HIV Ag/Ab Combo assay (chemiluminescence Immunoassay), and the viral load results of HIV-1 nucleic acid testing (NAT) were both>5 000 copies/ml. The patient can be diagnosed with HIV infection according to the National Guideline for Detection of HIV/AIDS(2020 Revised Edition). The thorough investigation findings and supplementary experiment results indicated that the false-positive HIV-1 NAT results was caused by cross-reactivity between the target sites detected by conventional HIV-1 NAT reagents and the lentiviral vectors fragments integrated into the genome of patient′s hematopoietic stem/progenitor cells. In conclusion, it is important for laboratories to select appropriate HIV-1 NAT testing platforms which won′t cause cross-reactivity for the testing of samples from patients who have been treated with HIV-derived vectors. It is also recommended to design and develop NAT testing platforms with multiple target regions labeled by different fluorescents for HIV NAT supplementation experiment to reduce the risk of false-positive diagnoses of HIV infection.

Effect of the transcription factor SOX3 on ovarian granulosa cell proliferation and estradiol secretion / 安徽医科大学学报

Objective @#To study the effect of sex-determining region Y-frame protein 3 (SOX3) on proliferation and estradiol secretion in human ovarian granulosa cells (KGN cell line) . @*Methods@#The gene sequence of human SOX3 (NM_005634. 3) was searched in Gene-Bank , an NCBI database , and the target gene SOX3 was amplified by PCR , which was cloned into lentiviral vector pLV-EF1a-GFP-2A-Puro to obtain the overexpression lentiviral re- combinant plasmid pLV-EF1a-GFP-2A-Puro- SOX3 ; the correctly sequenced overexpressed lentiviral recombinant plasmid as well as packaging plasmids ( pGag/Pol , pRev , pVSV-G) were co-transfected into human embryonic kidney cell line ( HEK 293T) cells ( pLV-SOX3 group) , and pLV-EF1a-GFP-2A-Puro and packaging plasmids (pGag/Pol , pRev , pVSV- G) were co-transfected into HEK 293T cells (pLV-NC group) , the lentiviral particles of both groups were collected and the titers of the viruses were measured after 48 h of transfection , the lentiviruses of the two groups were infected into KGN cells , and the stably expressed cell lines were obtained after puromycin screening for 2 weeks; real-time fluorescence quantitative PCR (RT-qPCR) and Western blot were used to detect the SOX3 mRNA and protein levels in the two groups; CCK-8 assay was used to detect the proliferative ability of the cells in the two groups; ELISA was used to determine the concentration of estradiol in the two groups .@*Results@#The identification of PCR products and sequencing results showed that the SOX3 gene fragment was amplified successfully , and the enzyme digestion and sequencing results indicated that the construction of overexpression lentiviral recombinant plasmid was completed; green fluorescence could be detected after lentiviral infection of HEK 293T cells , which indicated that lentiviral packaging was successful; the lentivirus was screened by puromycin after lentiviral infection of KGN cells , and the cells were observed to express green fluorescence under the fluorescence microscope; RT- qPCR and Western blot assays both showed that the expression level of SOX3 in the pLV-SOX3 group was significantly higher than that in the pLV-NC group ( P < 0. 05) . CCK-8 assay results showed that the proliferation ability of the cells in the pLV-SOX3 group significantly increased compared with that in the pLV-NC group (P < 0. 01) . ELISA results showed that estradiol concentration was elevated in the pLV-SOX3 group com- pared with the pLV-NC group (P < 0. 05) . @*Conclusion@#Overexpression of the transcription factor SOX3 can pro- mote the proliferation and estradiol secretion of human ovarian granulosa cells KGN .

Development,verification and application of fluorescence quantitative PCR method for detection of virus titer of recombinant measles virus vector SARS-CoV-2 vaccine / 中国生物制品学杂志

@#Objective To develop,verify and preliminarily apply a fluorescence quantitative PCR(qPCR)method for detection of the virus titer of recombinant measles virus vector severe acute respiratory symptom coronavirus 2(SARS-CoV-2)vaccine.Methods SARS-CoV-2 S gene was amplified using recombinant plasmid pUC57-S351 as the template,and the primer concentration was optimized to develop the qPCR detection method.The specificity and repeatability of the method were verified,and the linear range and limit of detection were determined.The copy number of recombinant virus S gene was detected by the developed qPCR method 1～12 d after continuous culture in bioreactor.Results The qPCR method was developed with the primer concentration of 0.20 μmol/L,which specifically detected the copy number of SARS-CoV-2 S gene.The linear relationship was good(R2= 0.995)at the template concentration ranged from 2 × 10~2 to 2 × 10~8 copies/μL,and the limit of detection was 2 × 10~2 copies/μL;The coefficient of variation(CV)value of 6 repeated detections of copy number of recombinant virus S gene was 2.64%.The copy number of recombinant virus S gene was monitored by the developed qPCR method 1 ～ 12 d after continuous culture in bioreactor,and the results were basically consistent with those detected by cytopathic method.Conclusion The developed qPCR method has good specificity and repeatability,which can be used for virus titer detection of recombinant measles virus vector SARS-CoV-2 vaccine.

Construction and expression of lentiviral vector of p62 gene over expression / 安徽医科大学学报

Objective @#To construct lentiviral vector of p62 gene over expression , and stably express p62 gene in human monocytic leukemia cells 1 (THP 1) , and to provide a way to study the role of p62 gene at the cellular lev el .@*Methods @#The p62 gene fragment was amplified by polymerase chain reaction (PCR) , and the amplified product was ligated to the linearized pcDNA3 1 Flag PCDH10 lentiviral vector. After identifying with PCR , the PCR product was cotransfected with the packaging plasmid into human embryonic kidney cells 293 (HEK 293T) . THP 1 cells were infected with recombinant lentivirus . Positive cell clones were screened by ampicillin . Western blot and real time fluorescence quantitative polymerase chain reaction (RT qPCR) were used to detect THP 1 cell lines with high p62 expression ( overexpression group) and THP 1 cell lines transfected with empty plasmid without p62 gene ( control group) . The expression levels of TNF α, IL 1βand Cxcl1 after K. p. infection were detected by RT qPCR .@*Results @#The p62 gene fragment was successfully obtained by PCR and ligated to pcDNA3 1 Flag PCDH10 vector. PCR confirmed that p62 pcDNA3 1 Flag PCDH10 recombinant plasmid was constructed successfully. Am picillin resistant cell lines were selected after lentiviral infection of THP 1 cells . The results of Western blot analysis showed that the THP 1 cells with drug sieve survival increased the expression of P62 protein compared with the con trol cells (P < 0.001) , and RT qPCR analysis showed that the relative mRNA expression of p62 increased (P < 0.001) . THP 1 cells with high expression of P62 were successfully constructed . The levels of TNF-α、IL-1βand C xcl1 from THP 1 cells with high expression of P62 significantly increased after infection with K. p. (P < 0.01) . @*Conclusion @#P62 pcDNA3 1 Flag PCDH10 vector and THP 1 cells with high expression of P62 can be successful ly constructed by three plasmid packaging system , which provides a basis for the study of p62 .

Vector analysis outcomes after Femtosecond Laser In-Situ Keratomileusis (FS-LASIK) versus Small Incision Lenticule Extraction (SMILE) for moderate myopic astigmatism

Objectives@#To compare the vector analysis, visual, and refractive outcomes of femtosecond-assisted laser insitu keratomileusis (LASIK) and small incision lenticule extraction (SMILE) among myopic patients with moderate myopic astigmatism.@*Methods@#This was a single-center, retrospective, cohort study that compared eyes that underwent femtosecond LASIK or SMILE for the correction of myopia and astigmatism of 0.75 to 3.0 diopters. Vector analysis and standard graphs for reporting visual and refractive outcomes were utilized for analysis.@*Results@#There were 82 femtosecond LASIK-treated eyes and 80 SMILE-treated eyes with similar preoperative characteristics except for slightly higher mean preoperative sphere refraction in the SMILE group (-4.2±2.4 D vs -4.9±1.6 D, p=0.03). At 3 months, femtosecond LASIK group had better mean uncorrected distance visual acuity (UDVA) (LogMAR 0.006±0.06 vs 0.06±0.09, p=0.00) and had more eyes achieving postoperative UDVA of 20/20 or better (88% versus 56%). Although there were similar postoperative spherical equivalents, residual astigmatism was higher in the SMILE group (0.11±0.22 D vs 0.32±0.30 D, p=0.00). Vector analyses showed significantly better outcomes for femtosecond LASIK than for SMILE in terms of difference vector (DV), index of success (IOS), torque, and flattening index (FI). A trend for undercorrection for higher astigmatism was seen in both groups that was greater in the SMILE group. Both groups showed high safety with the majority of eyes showing postoperative corrected distance VA (CDVA) within 1 line of preoperative CDVA (98.8% versus 91.2%). @*Conclusion@#Although femtosecond LASIK and SMILE have similar predictability at 3 months, femtosecond LASIK has relatively better efficacy and superior astigmatic outcomes than SMILE for the correction of moderate myopic astigmatism.

Biological threats to global malaria elimination Ⅲ Vector insecticide resistance / 中国血吸虫病防治杂志

The insecticide resistance is becoming increasingly severe in malaria vectors and has become one of the most important threats to global malaria elimination. Currently, malaria vectors not only have developed high resistance to conventional insecticides, including organochlorine, organophosphates, carbamates, and pyrethroids, but also have been resistant to recently used neonicotinoids and pyrrole insecticides. This article describes the current status of global insecticide resistance in malaria vectors and global insecticide resistance management strategies, analyzes the possible major challenges in the insecticide resistance management, and proposes the response actions, so as to provide insights into global insecticide resistance management and contributions to global malaria elimination.