Development and verification of cation exchange high performance liquid chromatography for detection of charge variants of pembrolizumab / 中国生物制品学杂志

@#Objective To develop and verify a cation exchange high performance liquid chromatography(CEX-HPLC)method for the detection of charge variants of pembrolizumab.Methods Pembrolizumab was bound to the exchange column matrix by using MabPac SCX-10 column,and the variants with different charges were eluted by gradually increasing the salt concentration of the mobile phase.The specificity,precision,linear range,accuracy and durability of the method were verified,and the charge variants of three batches of pembrolizumab finished products were detected by using the developed method.Results The resolution of the last acidic isomer peak and the first basic isomer peak of pembrolizumab from the main peak were 1.28 and 1.42,respectively.The mobile phase A and preparation buffer had no obvious interference peaks at the peak of the sample;The RSD values of the precision verification were all less than 2.0%;The total peak area,main peak area,acidic isomer peak area and basic isomer peak area of the standard all exhibited good linear relationship with the theoretical dilution concentration with each R~2 of 1.00;The recovery rates of the total peak area and main peak area of the standard at three concentrations were between 96.81% and 106.07%;When pH value of the mobile phase was within the range of 6.30±0.10,the RSD values of the total peak area and main peak area percentage of the standard were1.5% and 1.9%,and when the column temperature was within the range of(35±4) ℃,the RSD values of the total peak area and main peak area percentage of the standard were 0.4% and 0.3%,respectively.The RSD value of the main peak retention time of the three batches of finished products was 0.Conclusion The developed CEX-HPLC method can effectively separate the acidic isomers,main peaks and basic isomers of pembrolizumab with good specificity,precision and accuracy,which can be used for the follow-up research and development of pembrolizumab,the process verification of expanding production and the stability research.

Role of Macrophage Polarization in Chronic Low-grade Inflammation of PCOS Based on Theory of Spleen Being in Charge of Defensive Function / 中国实验方剂学杂志

Polycystic ovary syndrome （PCOS） is a reproductive endocrine disorder characterized by coexisting reproductive dysfunction and glucolipid metabolic disturbance， affecting 8%-13% of women of reproductive age and 3%-11% of adolescent females. Due to the highly heterogeneous clinical features， symptom-oriented individualized strategies are commonly adopted for the treatment of PCOS. Chronic low-grade inflammation is one of the core mechanisms for the occurrence of PCOS. Macrophages， as foundational cells of innate immunity， play an indispensable role in modulating systemic inflammatory responses. The imbalance of macrophage M1/M2 polarization is involved in chronic low-grade inflammation in PCOS via pathways such as activating pro-inflammatory responses， disrupting ovarian tissue repair， stimulating excessive synthesis of androgens， and promoting the occurrence of insulin resistance. Reshaping the phenotype of macrophages might serve as a potential therapeutic strategy for PCOS. Traditional Chinese medicine （TCM） holds that spleen deficiency and phlegm dampness is a crucial pathogenesis of PCOS. The spleen， being in charge of defensive function， plays a key role in ensuring normal physiological functions such as transportation and defense against external pathogen during the occurrence and development of PCOS. The imbalance of macrophage polarization resembles the transition from spleen being in charge of defensive function to spleen losing its defensive role in TCM. Therefore， this paper， for the first time， explores the deep connection between macrophage polarization and the pathogenesis of chronic low-grade inflammation in PCOS from the TCM theory of spleen being in charge of defensive function， providing theoretical support and new research directions for the treatment and drug research of PCOS.

Uma clínica em extensão: a psicanálise na atenção básica / Une clinique d'extension: la psychanalyse dans les soins primaires / An extension clinic: psychoanalysis in primary care

Nas últimas décadas, observamos que a psicanálise expandiu suas fronteiras para além dos seus espaços tradicionais, como em núcleos de ensino e pesquisa universitários e serviços e movimentos psiquiátricos e de saúde mental, dividindo-se em várias correntes e em diversas associações e escolas psicanalíticas. O percurso da disciplina psicanalítica nos mostra que vários desafios foram impostos à psicanálise, dentre estes a necessidade de problematizar os campos de intervenção possíveis. O psicanalista que atua no campo institucional, a partir do seu trabalho com pacientes e equipes de saúde, deve se responsabilizar pela transmissão do saber que ali se constrói, visto que essa clínica, ao ocorrer na instituição de saúde, produz efeitos tanto para a clínica quanto para a própria instituição. Portanto, este trabalho tem por objetivo apresentar, a partir das vinhetas clínicas, as intervenções possíveis do psicanalista junto às equipes de saúde da Atenção Básica sob a perspectiva do matriciamento

Au cours des dernières décennies, nous avons observé que la psychanalyse a élargi ses frontières au-delà de ses espaces traditionnels, comme dans les centres d'enseignement et de recherche universitaire et dans les services et mouvements psychiatriques et de santé mentale, se divisant en plusieurs courants et diverses associations et écoles psychanalytiques. Le parcours de la discipline psychanalytique nous montre que plusieurs défis se sont imposés à la psychanalyse,parmi lesquels la nécessité de problématiser les champs d'intervention possibles. Le psychanalyste qui travaille dans le champ institutionnel, à partir de son travail avec les patients et les équipes soignantes, doit avoir la responsabilité de transmettre les connaissances qui s'y construisent, puisque cette clinique, lorsqu'elle se déroule dans l'établissement de santé, produit des effets tant pour la clinique que pour ainsi que pour l'institution elle-même. Ce travail vise donc présenter, à partir de vignettes cliniques, les interventions possibles du psychanalyste auprès des équipes de soins de première ligne dans une perspective d'accompagnement matriciel

In recent decades, we have observed that psychoanalysis has expanded its borders beyond its traditional spaces, such as in university teaching and research centers and psychiatric and mental health services and movements, dividing into several currents and various psychoanalytic associations and schools. The path of the psychoanalytic discipline shows us that several challenges were imposed on psychoanalysis, among them the need to problematize the possible fields of intervention. The psychoanalyst who works in the institutional field, based on his work with patients and healthcare teams, must be responsible for transmitting the knowledge that is constructed there, since this clinic, when taking place in the healthcare institution, produces effects both for the clinic as well as for the institution itself. Therefore, this work aims to present, based on clinical vignettes, the psychoanalyst's possible interventions with Primary Care health teams from the perspective of matrix support.

Quality evaluation of anti-PD-1/CTLA-4 bispecific antibody / 中国生物制品学杂志

@#Objective Establish quality control methods for critical quality attribute of bispecific antibody against programmed cell death protein 1(PD-1)/cytotoxic T-lymphocyte-associated protein 4(CTLA-4).Methods The biological activity of PD-1 target was determined by reporter gene assay,and the competitive binding activity of CTLA-4 target was determined by flow cytometry;The antibody molecular size variants were controlled by reducing/non-reducing capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS) and size exclusion chromatography-high performance liquid chromatography(SEC-HPLC);Charge heterogeneity was determined by imaging capillary isoelectric focusing electrophoresis(iCIEF);Bispecific anti-PD-1/CTLA-4 antibody was identified by peptide map analysis;Glycosylation was analysed by high performance liquid chromatography(HPLC)Results The concentration for 50% of maximal effect(EC_(50)) of PD-1target was(6.91±0.78) nmol/L,and the relative biological potency to the reference was(103.50±13.08)% with the RSD of 12.64%;The EC_(50) of CTLA-4 target activity was(0.35±0.28) nmol/L,and the relative biological potency was(99.30±9.15)% with the RSD of 8.32%.The percentage of peak area of light chain and heavy chain of reducing CE-SDS was(98.86±0.02)%.The main peak area percentage of non-reducing CE-SDS was(93.07±0.13)%,fragment percentage was(4.44±0.13)%,and polymer percentage was(2.49±0.15)%.The peak area percentage of SEC-HPLC monomer and polymer were(97.20±0.01)% and(2.68±0.01)%,respectively.The area percentage of peak A group,peak B group,peak C group and peak D group were(38.43±0.54)%,(43.26±0.32)%,(11.31±0.14)% and(7.00±0.17)%,respectively.Peptide mapping showed the specific spectrum of the bispecific anti-PD-1/CTLA-4 antibody,which could be adopted for identification test.The highest proportion of glycotype was GOF,with a content of(41.06±0.11)%,There were three types of glycan containing sialic acid,namely G2F+G1F-NANA,G2F-NANA and G2F-2NANA,with the content of(12.44±0.12)%,(12.00±0.05)% and(5.37±0.05)%,respectively.The total content of glycan containing sialic acid was(29.80±0.20)%.Conclusion The critical quality attributes of bispecific anti-PD-1/CTLA-4 antibody were studied and the corresponding quality control methods were established to ensure its safety,effectiveness and quality control,which provides a reference for the quality control methods and strategies of this type of monoclonal antibody products.

Development and verification of whole-column imaging capillary isoelectric focusing electrophoresis analysis method for charge heterogeneity of human growth hormone Fc fusion protein / 中国生物制品学杂志

@#Objective To develop and verify a whole-column image capillary isoelectric focusing(iCIEF) electrophoresis method to analyze the charge heterogeneity of recombinant human growth hormone Fc fusion protein(Fc-rhGH).Methods The iCIEF analysis method of Fc-rhGH was developed by optimizing the target protein concentration,cosolvent(urea)concentration and focusing time.The target protein was simultaneously analyzed by this method and traditional flat plate isoelectric focusing(IEF) electrophoresis,and the results were compared;The specificity,accuracy,precision,limit of quantitation(LOQ) and durability of the developed method were verified.Results The optimized method was using the mixed solution of 8 mol/L urea,0.35% methyl cellulose(MC),4% amphoteric electrolyte and 0.5% isoelectric point marker as the sample buffer,and the focusing condition was 1 500 V 1 min,3 000 V 5.5 min.IEF was not suitable for analyzing the charge heterogeneity of Fc-rhGH solution.Using the optimized iCIEF for analysis,the target protein was significantly different from the unrelated protein,and the baseline of blank reagent was stable;The recovery rate of accuracy verification was within 90%～110%,and the linear range was 0.25～0.75 mg/mL(50%～150% of the target loading volume);The RSD of each isomer pI in the repeatability verification was less than 0.3%,and the RSD of peak area percentage was less than 5%;The LOQ was 0.04 mg/ml.The sample storage time durability,amphoteric electrolyte pharmalyte 3-10 durability and MC durability of this method were good.Using this method to analyze the charge heterogeneity of Fc-rhGH physicochemical reference substance,eight charge heterogeneities of the reference substance were effectively separated,and the pI ranged from 5.9 to 6.4.Conclusion The developed iCIEF method had good specificity,accuracy,precision and durability,and was more suitable for efficient analysis of charge heterogeneity of Fc-rhGH than traditional flat plate IEF,which was of great significance for the quality control of Fc-rhGH and other Fc fusion proteins.

Immune effect of electric charge modified influenza vaccine liposomes / 中国生物制品学杂志

@#Objective To explore the physical and chemical properties of electric charge modified liposomes and the immune enhancement effect as influenza vaccine adjuvants.Methods Four kinds of electric charge-modified liposomes were prepared with soybean lecithin,cholesterol,N-1-(2,3-dioleyloxy) propyl-N,N,N-trimethyllam-moniumchloride(DOTAP)and 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC)(DOTAP and DOPC mass ratio 1:4,2:3,3:2 and 4:1).The tetravalent influenza vaccine was mixed with the four liposomes according to the volume fraction of 1.4:1.0,which was prepared into cationic modified influenza vaccine liposome freeze-dried powder by freeze-thaw freeze-drying method.The liposome freeze-dried powder was redissolved with PBS,the particle size and Zeta potential were measured by laserparticle size analyzer,and the encapsulation efficiency of liposome was measured by Lowry method.231 female KM mice were randomly divided into negative control group(PBS),positive control group(tetravalent vaccine bulk),cationic liposome group(four electric charge modified influenza vaccine liposomes) and neutral liposome group(non-electric charge modified influenza vaccine liposomes),which were injected intraperitoneally at 7,14 and 28 d,0.2 mL per mouse.The cellular immune effect was evaluated by MTT assay and T lymphatic surface labeling,and the humoral immune effect was evaluated by hemagglutinin inhibition(HI) at 3,7,14,28,42 and 56 d of immunization.Results When the mass ratio of DOTAP to DOPC was 4:1,the particle size and Zeta potential of cationic modified liposomes reached the maximum,which were 529.65 nm and 12.05 mV respectively,and the encapsulation efficiency was also high,which was 81.82%.Stimulus index(SI) and CD4~+/CD8~+value of spleen cells of mice in the four cationic liposome groups were significantly higher than those in the negative control group(F=4.651～25.866,each P<0.05),among which,the two indexes in 4:1 cationic liposome group were higher than those in the other three groups;mice in this group produced early humoral immune effect 3 d after immunization,and the antibody titer reached the highest 42 d after immunization and maintained a high level during the whole immunization cycle.Conclusion Cationic liposomes with different electric charge modifications improved the immune effect of influenza vaccine,the immune enhancement effect was the best and the immune effect lasted the longest when the mass ratio of DOTAP to DOPC was 4:1.

Effects of simulated gas of warm pressure charge explosion on neurobehavior and hippocampal neurogenesis in rats / 中华行为医学与脑科学杂志

Objective:To investigate the effects of thermobaric charge explosion simulated gas on long-term neurobehavior and hippocampal neurogenesis in rats.Methods:A total of 48 male SPF grade SD rats aged 8-10 weeks were randomly divided into control group, 5 min exposure group, 10 min exposure group and 15 min exposure group, with 12 rats in each group. Twenty-eight days after inhalation of infection, the anxiety-like behavior of rats was evaluated by an elevated cross maze, and the learning and memory function of rats was evaluated by two-way active avoidance experiment. The number of positive cells of rat hippocampal dentate gyrus neural stem cells marker molecule neural epithelial cell protein (SOX2) and mature neuron marker molecular neuronal nuclei (NeuN) was detected by immunofluorescence staining. Western blot was used to detect SOX2 and NeuN protein expression in the hippocampal tissues of rats. GraphPad prism 8.0 software was used for data analysis.The comparison of repeated measurement design data was carried out by repeated measurement ANOVA.One-way ANOVA was used for inter group comparisons, and Tukey test was used for pairwise comparison. Hippocampal nerve cells were counted using the Image J software.Results:(1) The experimental results of the elevated cross maze showed that the percentage of arm opening and the percentage of open arm residence time in each group had significant group effects ( F=22.31, 5.43, all P<0.05). The percentage of open arm entry times of rats in the 5 min, 10 min and 15 min exposure group ((28.85±1.47)%, (15.04±4.69)%, (12.66±2.89)%) and the percentage of residence time in open arm ((12.12±2.64)%, (12.16±1.11)%, (8.73±3.52)%) were all lower than those of the control group ((65.40±1.86)%, (42.92±3.12)%) (all P<0.05). There were no statistically significant differences in pairwise comparison among the three exposure groups (all P>0.05). (2)During the memory acquisition period, the results of repeated-ANOVA showed that the time main effect ( F=56.46), the group main effect ( F=16.64) and the interaction effect had significant differences( F=4.21)(all P<0. 05). The difference values of active avoidance number between the 4th day and 1st day among the four groups were significant different ( F=68.63, P<0.05). During the memory reproduction period, there were significant differences in active avoidance number and active avoidance time among the four groups ( F=8.17, 8.28, both P<0.05). The active avoidance numbers in 10 min and 15 min exposure groups((2.50±0.26) times, (2.33±0.06) times)were significantly lower than those in the control group ((8.33±3.72) times) (both P<0.05), and the active avoidance time ((6.25±0.40)s, (6.61±1.63)s) were significantly higher than those in the control group((3.69±1.41)s) (both P<0.05). The active avoidance numbers in 10 min and 15 min exposure groups were significantly lower than that in 5 min exposure group (both P<0.05). (3) The results of immunofluorescence staining showed that the numbers of SOX2-positive cells in the four groups were statistically significant ( F=5.33, P<0.05). The SOX2-positive cells in 15 min exposure group (4.33±1.12) was significantly lower than that in control group (7.67±1.52) ( P<0.05). The numbers of NeuN-positive cells in the four groups were significantly different ( F=11.06, P<0.05), and the NeuN-positive cells in the 10 min and 15 min exposure groups((105.67±8.50), (88.33±9.50)) were significantly lower than that in the control group (127.00±6.56) ( P<0.05). The NeuN-positive cells in 15 min exposure group were significantly lower than that in 5 min exposure group (110.67±8.32) ( P<0.05). (4) Western blot results showed that the relative expression of SOX2 and NeuN proteins in the four groups was statistically significant ( F=11.560, 7.035, both P<0.05). The relative expression of SOX2 and NeuN proteins in the 15 min exposure group were significantly lower than those in control group (both P<0.05). The relative expression of SOX2 protein in 15 min exposure group was significantly lower than that in 5 min exposure group ( P<0.05). Conclusion:Acute exposure to warm pressure charge explosion simulated gas can lead to anxiety-like behavior, learning and memory deficits in rats, and significantly reduce the protein expression levels of hippocampal dentate gyrus neural stem cells and mature neuronal marker molecules SOX2 and NeuN.

Application of biological mass spectrometry in quality control of adeno-associated virus carrier preparations / 中国药科大学学报

@#Adeno-associated virus (AAV) is a common viral vector used in gene therapy.Because of its high safety and its ability to target a variety of cells, it has been widely used in preclinical and clinical studies.However, during the design and production, AAV vectors have many key quality attributes that affect their safety and efficacy.The development and application of biological mass spectrometry technology provides a convenient platform for the research on biological macromolecules, especially in the aspects of protein sequence, structure and interaction.For AAV vectors, mass spectrometry can facilitate the determination or characterization of capsid protein ratio, post-translational modification, serotype, and empty capsid ratio, thus assisting in the quality control of AAV vectors.Compared with the existing methods, mass spectrometry has the advantages of smaller amount of sample size, faster and more sensitive analysis, being more suitable for the analysis of complete AAV vectors with higher mass resolution, and can distinguish empty capsids, full capsids and partial capsids.In the future, mass spectrometry technology is expected to play a more important role in the design and production of AAV vectors through the coupling of more efficient protein separation technology with mass spectrometry, the development of new information processing software platforms and new mass spectrometry detection techniques.

Efficient biosynthesis of γ-aminobutyric acid by rationally engineering the catalytic pH range of a glutamate decarboxylase from Lactobacillus plantarum / 生物工程学报

γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase LpGAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of LpGAD based on surface charge. A triple point mutant LpGADS24R/D88R/Y309K was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and LpgadS24R/D88R/Y309K genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass (OD600) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of LpGAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.

A comprehensive review on photon-counting computed tomography: Principles, technical hurdles and analysis of clinical applications / 生物医学工程学杂志

In recent years, photon-counting computed tomography (PCD-CT) based on photon-counting detectors (PCDs) has become increasingly utilized in clinical practice. Compared with conventional CT, PCD-CT has the potential to achieve micron-level spatial resolution, lower radiation dose, negligible electronic noise, multi-energy imaging, and material identification, etc. This advancement facilitates the promotion of ultra-low dose scans in clinical scenarios, potentially detecting minimal and hidden lesions, thus significantly improving image quality. However, the current state of the art is limited and issues such as charge sharing, pulse pileup, K-escape and count rate drift remain unresolved. These issues could lead to a decrease in image resolution and energy resolution, while an increasing in image noise and ring artifact and so on. This article systematically reviewed the physical principles of PCD-CT, and outlined the structural differences between PCDs and energy integration detectors (EIDs), and the current challenges in the development of PCD-CT. In addition, the advantages and disadvantages of three detector materials were analysed. Then, the clinical benefits of PCD-CT were presented through the clinical application of PCD-CT in the three diseases with the highest mortality rate in China (cardiovascular disease, tumour and respiratory disease). The overall aim of the article is to comprehensively assist medical professionals in understanding the technological innovations and current technical limitations of PCD-CT, while highlighting the urgent problems that PCD-CT needs to address in the coming years.

Investigation of the influence of clathrate hydrate crystals on the structuring of homeopathic high dilutions

High dilutions (HD) of drugs used in homeopathy are mostly too dilute to contain original drug molecules. But evidences support their specific biological and therapeutic effects. The reason behind this is thought to be water structure characteristic of the original drug. Spectroscopic studies indicate that the specific water structure in HDs can be resolved into free water molecules, hydrogen bonding strength of water hydroxyl, number of hydrogen bonds and clathrate hydrate crystals (CHC). HDs are prepared in EtOH water solution by serial dilution and mechanical agitation, and are called potencies. The objective of the present study is to further confirm the presence of CHCs in the two potencies of three drugs. Electronic spectra of the HDs of the potencies indicate two broad peaks and marked difference in intensities of absorption. Furior Transform Infrared (FT-IR) spectra of the test potencies and their control show difference in intensity shift and contour shape of OH stretching and bending bands. All the experimental data indicate the presence of CHCs in varying amounts in the test potencies.

Acompanhamento do comportamento do HIV através de exames laboratoriais em portadora do vírus: relato de caso / Monitoring of HIV behavior through laboratory tests in a virus carrier: case report

O vírus da imunodeficiência humana é o agente etiológico da AIDS, doença crônica que destrói o sistema imunológico e é caracterizada pela baixa contagem de células TCD4, alta contagem de partículas virais no sangue e manifestações clínicas da doença. O diagnóstico se dá com o aparecimento de infecções oportunistas, que levam a contagem de TCD4 a níveis menores que 200 céls/mm³. Os exames laboratoriais para o diagnóstico do HIV foram os principais avanços para o início do tratamento, reduzindo a transmissão. Detecção de anticorpos, detecção de antígenos e amplificação do genoma do vírus são alguns dos exames laboratoriais utilizados para diagnóstico. Os dois principais biomarcadores são os exames de contagem de células TCD4, que verifica o sistema imune, e a quantificação de carga viral, que informa a quantidade de partículas virais, mostrando a progressão da infecção. Quanto maior a carga viral, maior o dano ao sistema imune. Uma carga viral indetectável é inferior a 50 cópias/mL, mas valores menores ou iguais a 200 cópias/mL também impedem a transmissão. Uma declaração de consenso afirma que Indetectável é igual a Intransmissível. Portanto, quando indetectável, a transmissão inexiste. O presente estudo relata e discute o caso clínico de uma paciente diagnosticada com HIV/AIDS aos 28 anos, que sobreviveu, apesar do diagnóstico tardio, e sob presença de doença oportunista com um grave grau de diminuição de células TCD4 (22 cél/mm³). Por meio do diagnóstico, introdução e adesão correta da terapia antirretroviral e monitorização de exames laboratoriais, conseguiu evitar a morte e ter uma vida semelhante à de um HIV negativo. Ultrapassou a expectativa de vida que na descoberta era de 10 anos, com uma qualidade de vida considerável, não sendo transmissora do vírus, diminuindo assim o estigma e preconceito. O biomédico é peça fundamental nesse contexto, considerando que deve fornecer informações precisas e fidedignas, tão necessárias ao acompanhamento de pessoas vivendo com HIV, para que autoridades e profissionais de saúde adotem medidas adequadas, tanto na prevenção, quanto no diagnóstico e monitoramento da doença.

The human immunodeficiency virus is the etiological agent of AIDS, a chronic disease that destroys the immune system and is characterized by low TCD4 cell count, high viral particle count in blood and clinical manifestations of the disease. The diagnosis is due to the appearance of opportunistic infections, which lead to TCD4 counts below 200 cells / mm³. Laboratory tests for the diagnosis of HIV were the main advances in starting treatment, reducing transmission. Antibody detection, antigen detection and virus genome amplification are some of the laboratory tests used for diagnosis. The two main biomarkers are the TCD4 cell count tests, which checks the immune system, and viral load quantification, which reports the number of viral particles, showing the progression of infection. The higher the viral load, the greater the damage to the immune system. An undetectable viral load is less than 50 copies / mL, but values less than or equal to 200 copies / mL also prevent transmission. A consensus statement states that Undetectable equals Non-Transmissible. Therefore, when undetectable, transmission does not exist. The present study reports and discusses the clinical case of a patient diagnosed with HIV / AIDS at age 28, who survived despite late diagnosis and under the presence of opportunistic disease with a severe degree of TCD4 cell reduction (22 cells / mm³). Through the diagnosis, introduction and correct adherence of antiretroviral therapy and monitoring of laboratory tests, she was able to avoid death and have a life similar to that of an HIV negative. Exceeded the life expectancy that in the discovery was 10 years, with a considerable quality of life, not transmitting the virus, thus reducing the stigma and prejudice. The biomedical is a key player in this context, considering that he must provide accurate and reliable information, which is so necessary for the monitoring of people living with HIV, so that authorities and health professionals adopt appropriate measures, both in prevention, diagnosis and monitoring of the disease.

High Dilutions of Drugs show Distinct Variation from each other in their Electronic Spectra

Drugs at high dilution (HD) produce therapeutic effect on man, animals and plants. Experimental evidence shows that free water molecules and hydrogen bond strength of OH groups constitute the physical basis of HDs which are otherwise devoid of original drug molecules. HDs are produced in aqueous EtOH by serial dilution of a substance with mechanical agitation or succussion in each step, and are called potencies. Three potencies 6 cH, 12 cH and 30 cH of two drugs Anacardium orientale and Natrum muriaticum(NaCl) and their mother tincture (MT) are used in this study. Electronic spectra of these MTs and potencies, all in 90% EtOH, were taken in the wavelength region of 190 nm ­350 nm. The objective is to find out any additional physical-chemical entities in potencies besides the aforesaid two factors. It was reported earlier that charge transfer (CT) interaction accompanies potentization of drugs. This study focused on the CT interaction. The results indicate that spectral pattern and absorbance intensities of the test samples vary from each other. Natm 6cH (absorbance 0.30 at 196.53nm), 12cH (abs. 0.06 at 196.53nm) and 30cH (abs. 1.32 at 196.5nm). Anac 6cH (abs. 0.33 at 203nm), 12cH (abs. 0.61 at 208nm) and 30cH (abs. 0.09 at 200.67nm). The spectrum of each potency shows two peaks. The 2nd peak at higher wave length belongs to CT interaction. Anac 6cH suc, 7cH unsuc. Insersections at 197.14nm with abs. 0.05, and 290nm with abs. 0.01. Anac 12cH suc, 13cH unsuc. Intersections at 196.93nm with abs. 0.06, and 273nm with abs. 0.00. Anac 30cH suc, 31cH unsuc. Intersections at 194.42nm with abs. -0.05, 238.03nm with abs. -0.01, 252.15nm with abs. -0.002, and 261nm with abs. 0.004. Natm 6cH suc, 7cH unsuc. Intersection at 199.44nm with Abs -0.11. Natm 12cH suc, 13cH unsuc. Instersection at 200.48nm with abs. -0.11. Natm 30cH suc, 31cH unsuc. Intersection at 204.24nm with abs. -0.08. Potentization involves CT interaction in consecutive potencies. Water and EtOH do not form a homogeneous mixture and have aggregates of EtOHand water molecules. CT interactions occur in these individual aggregates and are mostly inter molecular within EtOH or water. These aggregates vary from each other in the test samples. The spectra of test samples were analysed for margin of error (MOE). The MOE is very small (0.001-0.002%), and for this reason the difference between the spectra is significant. Besides that the intersection between consecutive spectra vary in number and position. It is concluded that water and EtOH aggregates and their relative distribution constitute additional physical-chemical basis of potencies.

Bilateral choanal atresia – An unusual case of severe respiratory distress in a newborn without nasal flaring

Respiratory distress in a newborn can be due to various causes and some need active intervention. Choanal atresia (CA) is a rare congenital anomaly with its incidence estimated to be 1 case/5,000–8,000 births. It is characterized by narrowing or blockage of the nasal passages. It is important to make an early clinical diagnosis with emphasis on timely management as it can be life-threatening. The pediatrician may be not able to pass a feeding tube through the neonate’s nostril even on repeated attempts. Detailed evaluation should be performed for the CHARGE association. High-resolution computed tomography can aid the diagnosis and transnasal endoscopic surgery is the preferred treatment modality. Here is a case report of a term neonate born with severe respiratory distress who was diagnosed to have bilateral CA on evaluation and managed with nasal endoscopic surgery.

Analysis of common problems in hospital medical service charge management / 中华医院管理杂志

As suggested by regulatory feedbacks from relevant national regulatory agencies on the use of medical insurance funds, noticeable problems were found in the compliance and rationality of medical service charges among Chinese hospitals. Based on the practice of hospital management, the authors summarized common problems of various medical service charge management as reported by the management authorities at all levels. These problems referred to name-replaceable charges and affiliated charges, splitting charges, repeated charges, and multiple(false) charges. On this basis, the paper analyzed such hospital management risks incurred as regulatory penalties, fee disputes and medical disputes, as well as their internal and external causes. In the end, the paper put forward countermeasures and suggestions for reference by hospitals in their management of medical service charges.

CHARGE syndrome in a neonate with esophageal atresia / 中华围产医学杂志

This article reported a case of neonatal CHARGE syndrome complicated by congenital esophageal atresia. A prenatal ultrasound examination at 30 weeks of gestation revealed polyhydramnios and a small magenblase of the fetus, then fetal MRI suggested congenital esophageal atresia. The infant was born with severe asphyxia at 37 +5 gestational weeks by cesarean section due to placental abruption with a birth weight of 2 310 g. Gastric tube could not be placed after resuscitation. Congenital esophageal atresia complicated by tracheoesophageal fistula was diagnosed by esophageal imaging. Bilateral choanal atresia was detected by electronic nasopharyngoscopy and MRI. Moreover, skull defect, suspected meningocele were also observed. CHARGE syndrome was confirmed by whole exome sequencing, revealing a frameshift deletion of c.2155delA (p.Thr719GlnfsTer9) in the CHD7 gene. The infant died after withdrawing treatment.

Relação entre o bem-estar subjetivo e o autocuidado em cuidadores em tempos pandêmicos / Relationship between subjective well-being and self-care among caregivers in pandemic times

O autocuidado se refere à práticas de prevenção de doenças e de manutenção da saúde. A capacidade de autocuidado pode ser um fator importante em diferentes condições e contextos, e associados aos aspectos da saúde do indivíduo podem proporcionar benefícios diretos a esta pessoa. Durante a Pandemia (COVID-19) esta capacidade pode estar diminuída ou prejudicada, e ainda imbricado com aspectos afetivos, de forma a influenciar o seu bem-estar subjetivo, ou seja, afetar sua avaliação do próprio bem-estar. Assim, esse Estudo teve como Objetivo avaliar a relação entre capacidade de autocuidado e a atribuição afetiva (positiva ou negativa) de adultos que atuam como cuidadores, dos gêneros masculino e feminino. Os instrumentos utilizados foram um questionário sociodemográfico, a Escala de Afetos Positivos e Negativos e a Escala para Avaliar as Capacidades de Autocuidado. Após a análise de frequência das variáveis, passou-se a análise de correlação de Pearson e por meio dela foram reveladas associações moderadas (médio altas) entre a medida de afeto positivo e autocuidado (r=0,62; p=0,000) e afeto negativo e autocuidado, neste caso, com coeficiente negativo (r=-0,42; p=0,000), indicando assim, que pessoas tendem a ter mais autocuidado, apresentar melhor afeto positivo e menor afeto negativo, respectivamente. Pôde-se inferir a ideia de que bem-estar e qualidade de vida estão imbricados à atribuição afetiva do indivíduo.

Self-care refers to disease prevention and health maintenance practices. Self-care capacity can be an important factor in different conditions and contexts and when associated with aspects of an individual's health it can provide direct benefits to this person. During the Pandemic (COVID-19) this ability may be diminished or impaired, and even hindered with affective aspects, influencing their subjective well-being, that is, affecting their assessment of their own well-being. Thus, this study aimed to evaluate the relationship between self-care capacity and the affective attribution (positive or negative) of male and female adults who act as caregivers. The instruments used were a sociodemographic questionnaire, the Positive and Negative Affect Schedule and the Scale to Assess Self-Care Capabilities. After analyzing the frequency of the variables, Pearson's correlation analysis was performed and through it, moderate associations (medium high) were revealed between the measure of positive affect and self-care (r=0.62; p=0.000) and negative affect and self-care, in this case, with a negative coefficient (r=-0.42; p=0.000); thus, this indicates that people tend to have more self-care, have better positive affects and less negative affects, respectively. It was possible to infer the idea that well-being and quality of life are intertwined with the affective attribution of the individual.

Os impasses ao acesso à intersubjetividade: observações clínicas de um bebê com risco de evolução autística / Impases en el accesoa la intersubjetividad: observaciones clínicas de un bebé con riesgo de evolución autista / Challenge in accessing intersubjectivity: clinical observations of a baby with a risk of autism / Les impasses à l'accès à l'intersubjectivité: observations cliniques d'un bébé à risque d'autisme

O artigo aborda os impasses vividos por um bebê com risco de autismo em seu processo de subjetivação. Estudos afirmam que há bebês que, muito precocemente, recusam-se a estabelecer trocas com o objeto materno, impossibilitando que a mãe entre em um estado de identificação com seu bebê, o que colocaria em pauta que o autismo infantil não se limitaria apenas a uma falha materna, mas se trataria de uma patologia multifatorial. Quais intervenções clínicas seriam necessárias para que a mãe e o bebê possam juntos (re)escrever uma nova narrativa psíquica? Haveria um tipo de intervenção e de manejo clínico específico nos casos de bebês com risco de autismo? E como a psicanálise contemporânea, fundamentada pela leitura da intersubjetividade, nos ajudaria na compreensão da clínica da intervenção precoce?

Este artículo se propone abordar los impases experimentados por un bebé con riesgo autista en su proceso de subjetivación. Existen estudios sobre bebés que precozmente se niegan a establecer intercambios con el objeto materno, imposibilitando que la madre entre en identificación con él, poniendo sobre la mesa que el autismo infantil no se limitaría a una falla materna, sino que se trataría de una patología multifactorial. Se plantea, cuáles intervenciones clínicas son necesarias para que madre y bebé puedan (re)escribir una nueva narrativa psíquica, que posibilite un verdadero encuentro diádico. ¿Habría un tipo específico de intervención y manejo clínico en los bebés con riesgo de autismo? ¿Cómo el psicoanálisis contemporáneo, basado en la lectura de la intersubjetividad y de lo intrapsíquico, ayudaría a entender la clínica de la intervención precoz?

This article addresses the failures of a baby with a risk of autism to develop his subjectivation process. Recent studies show that some babies are not able to develop exchanges with their maternal object, which would show that children autism would not be limited to a failure of the mother to enter into a state of identification with her baby, but would be associated to a multifactorial pathology. In that respect, which clinical interventions would be necessary so that mother and baby can (re)write a new psychic narrative, enabling them to develop a genuine capability to meet together? Is there a type of clinical handling for babies with a risk of autism? How can contemporary psychoanalysis, with focus on intersubjectivity and inter-psychism, could help us to understand early intervention clinic?

Cet article aborde les entraves d'un bébé à risque autistique à développer son processus de subjectivation. Des études récentes montrent que certains bébés ne sont pas capables de développer des échanges avec leur objet maternel, ce qui montrerait que l'autisme infantile ne se limiterait pas seulement à un échec de la mère d'entrer dans un état d'identification avec son bébé, mais qu'il s'agirait d'une pathologie multifactorielle. Ainsi, quelles interventions cliniques seraient nécessaires pour que mère et bébé puissent (ré)écrire ensemble un nouveau récit psychique, qui leur permettrait de développer une véritable capacité à se rencontrer ? Y aurait-il un type de prise en charge clinique spécifique aux bébés à risque d'autisme? Comment la psychanalyse contemporaine, centrée sur l'intersubjectivité et l'inter-psychisme, peut nous aider à comprendre la clinique d'intervention précoce?

Facteurs de risque et prise en charge de la rupture utérine dans une structure de 1ere référence du Mali: cas du district sanitaire de Bougouni / Risk factors and management of uterine rupture in a 1st reference structure in Mali: case of the Bougouni health district

L'objectif était d'évaluer les facteurs de risque de la RU et de proposer les aspects thérapeutiques. Matériels et méthodes : Nous avons réalisé une étude cas-témoins au centre de santé de Référence de Bougouni en 2019. Résultats : De janvier au 31 décembre 2019; sur 1161 accouchements 43 RU ont été enregistrées soit 3,7% correspondant à une RU pour 27 accouchements. Les patientes de 35 ans et plus ont été plus touchée par la RU (44,2%) avec ORaIC95%= 6,3 [1,5 - 26,3]. Les évacuations obstétricales avaient un ORaIC95%=25,6 [7,8- 83,7]. La totalité des patientes étaient des femmes au foyer (97,7%) des cas versus (82,3%) des témoins avec ORaIC95%= 8,9 (1,1-69). Les Paucipares et multipares avaient respectivement un ORaIC95%= 6,2 [1,8 - 20,3] et 4,1[1,3 - 12,9]. La cicatrice utérine (20,9%) des cas contre 8,1 % les témoins avait un ORaIC95%= 2,9 [1,1 - 8,7]. En effet l'absence de CPN étaient un facteur de risque, ORaIC95%= 3,0 [1,3 ­ 6,9]. Le délai de la RU était ˂ 6 heures chez 95%. En effet 34 RU complètes (79,1%) et 9 RU incomplètes (20,9%) ont été notées. Seulement 2,3 % des cas avaient accouché par voie basse. Le traitement de la RU reposait sur la chirurgie (100%) complétée par celui du choc (51,2%) des cas et de l'infection (100%) des cas. Conclusion: La RU est fréquente dans nos pays sous médicalisés. Sa prévention efficace passe par des stratégies visant à agir sur les facteurs de risque

The objective was to assess the risk factors for and to suggest therapeutic aspects. Materials and methods: We carried out a case-control study at the Bougouni Reference health center in 2019. Results: From January to December 31, 2019; out of 1161 deliveries, 43 uterine rupture were recorded, 3.7% corresponding to one uterine rupture for 27 deliveries. Patients 35 years and older were more affected by uterine rupture (44.2%) with ORaIC95% = 6.3 [1.5 - 26.3]. Obstetric evacuations had an ORaIC95% = 25.6 [7.8-83.7]. All of the patients were housewives (97.7%) versus (82.3%) controls with ORaIC95% = 8.9 (1.1-69). Pauciparous and multiparous had an ORaIC95% = 6.2 [1.8 - 20.3] and 4.1 [1.3 - 12.9], respectively. The uterine scar (20.9%) of cases versus 8.1% of controls had a 95% ORaIC95% = 2.9 [1.1 - 8.7]. Indeed the absence of ANC was a risk factor, ORaIC95% = 3.0 [1.3 - 6.9]. The time to uterine rupture was ˂ 6 hours in 95%. In fact 34 complete uterine rupture (79.1%) and 9 incomplete uterine rupture (20.9) were noted. Only 2.3% of cases gave birth vaginally. Treatment of uterine rupture was based on surgery (100%) supplemented by shock (51.2%) of cases and infection (100%) of cases. Conclusion: Uterine rupture is common in our countries under medical care. Its effective prevention involves strategies aimed at acting on risk factors.

Prévalence des hémorragies digestives au cours des vagues de COVID-19 au service de Réanimation Chirurgicale du CHU Joseph Ravoahangy Andrianavalona, Antananarivo, Madagascar / Prevalence of gastrointestinal bleeding during the waves of COVID -19 in the surgical intensive care unit of the CHU Joseph Ravoahangy Andrianavalona,Antananarivo, Madagascar

Introduction : La CoViD-19 est une maladie à « plusieurs visages ¼ qui peut affecter tous les systèmes. La survenue d'hémorragies digestives fait partie des manifestations de cette maladie. L'objectif de cette étude est de présenter les cas d'hémorragies digestives chez des patients infectés par le SARS-CoV-2. Matériels et Méthodes : Une étude rétrospective couvrant la période correspondant aux deux vagues de CoViD-19 à Antananarivo (Madagascar) a été réalisée, plus particulièrement au service de Réanimation Chirurgicale du Centre Hospitalier Universitaire Joseph Ravoahangy Andrianavalona. Résultats : Huit sur 101 patients de 51 à 81 ans, hospitalisés pour CoViD-19, ont présenté une hémorragie digestive dont les manifestations allaient de l'hématémèse au méléna ou une association de ces deux manifestations hémorragiques. Ces patients ont été traités par, entre autres une anticoagulation et une corticothérapie, comme défini dans le protocole national de prise en charge de la CoViD-19, avant l'épisode hémorragique. Aucun patient n'a présenté d'état de choc, l'indice de choc allait de 0,5 à 0,9. Deux patients ont pu bénéficier d'une fibroscopie digestive haute. Le score de Glasgow Blatchford variait de 6 à 13. Parmi ces huit patients, quatre sont décédés. Conclusion : Lors de la prise en charge de la CoViD-19, au vu des manifestations thrombotiques surtout, il faut procéder à une protection au niveau digestif lorsqu'une anticoagulation à titre curatif doit être réalisée. Également cette protection digestive doit être effectuée au-devant de la corticothérapie, laquelle entre dans le cadre du traitement de la CoViD-19. Tout cela pour minimiser le risque de survenue de saignement gastro-intestinal .

Background: CoViD-19 is a "many-faced" disease that can affect all the body organism. The occurrence of gastrointestinal bleeding is one of the manifestations of this disease. The aim of this study was to present cases of gastrointestinal bleeding in patients infected with SARS-CoV-2. Materials and Methods: A retrospective study covering the period corresponding to the two waves of CoViD-19 in Antananarivo (Madagascar) was carried out, more particularly in the Surgical Intensive Care Unit of the University Hospital Joseph Ravoahangy Andrianavalona. Results: Eight out of 101 patients aged 51 to 81, hospitalized for CoViD-19, presented gastrointestinal bleeding, with hematemesis or melena or a combination of these two bleeding manifestations. These patients were treated, among other coagulation and corticosteroid therapy as defined in the national protocol for the management of CoViD-19 before the bleeding episode. None of the patients presented with a shock; the shock index ranged from 0.5 to 0.9. Two patients were able to benefit from an upper digestive fibroscopy. The Glasgow Blatchford score ranged from 6 to 13. Of these eight patients, four died. Conclusion: During the management of CoViD-19, the thrombotic manifestations are treated with curative anticoagulation must be performed, which can cause digestive bleeding. Also, in front of the corticosteroid therapy which is part of the treatment of CoViD-19, also digestive protection must be carried out to minimize the risk of occurrence of gastrointestinal bleeding.