Treatment of periprosthetic joint infection after hip and knee arthroplasty / 中国组织工程研究

BACKGROUND:Periprosthetic joint infection is one of the most unwanted complications for surgeons and patients after arthroplasty,and its recalcitrance and intractability have always been a headache for arthroplasty surgeons. OBJECTIVE:To review the latest domestic and international clinical treatments used in the treatment of periprosthetic joint infection after hip and knee arthroplasty in recent years,including antibiotic treatment,surgical treatment,biological treatment and Chinese medicine treatment,to promote the research progress in the treatment of periprosthetic joint infection in China. METHODS:The literature from January 2000 to October 2022 on CNKI,WanFang,VIP,and PubMed was retrieved by the first author.762 articles were obtained by reading the titles for initial screening,then 194 articles were obtained by reading the abstracts and excluding studies with duplicate contents,low data reliability,and outdated views.Finally,88 articles were included through intensive reading of the original text. RESULTS AND CONCLUSION:(1)Combined antibiotic regimens may help eradicate the infection in the treatment of periprosthetic infections.(2)Two-stage revision remained the golden indicator for the treatment of periprosthetic infection.(3)One-stage revision lacked large-sample clinical studies and required more clinical observation.(4)Phage therapy and newer drug delivery systems in biological therapy had been applied in small amounts in the clinic,showing their advantages in the prevention and eradication of periprosthetic infections.(5)Chinese medicine with antibiotics and surgical treatment methods can improve the prevention and treatment of periprosthetic joint infection,but high-level evidence-based medical evidence was lacking.

Nanomaterial-based Therapeutics for Biofilm-generated Bacterial Infections / 生物化学与生物物理进展

Bacterial biofilms gave rise to persistent infections and multi-organ failure, thereby posing a serious threat to human health. Biofilms were formed by cross-linking of hydrophobic extracellular polymeric substances (EPS), such as proteins, polysaccharides, and eDNA, which were synthesized by bacteria themselves after adhesion and colonization on biological surfaces. They had the characteristics of dense structure, high adhesiveness and low drug permeability, and had been found in many human organs or tissues, such as the brain, heart, liver, spleen, lungs, kidneys, gastrointestinal tract, and skeleton. By releasing pro-inflammatory bacterial metabolites including endotoxins, exotoxins and interleukin, biofilms stimulated the body’s immune system to secrete inflammatory factors. These factors triggered local inflammation and chronic infections. Those were the key reason for the failure of traditional clinical drug therapy for infectious diseases.In order to cope with the increasingly severe drug-resistant infections, it was urgent to develop new therapeutic strategies for bacterial-biofilm eradication and anti-bacterial infections. Based on the nanoscale structure and biocompatible activity, nanobiomaterials had the advantages of specific targeting, intelligent delivery, high drug loading and low toxicity, which could realize efficient intervention and precise treatment of drug-resistant bacterial biofilms. This paper highlighted multiple strategies of biofilms eradication based on nanobiomaterials. For example, nanobiomaterials combined with EPS degrading enzymes could be used for targeted hydrolysis of bacterial biofilms, and effectively increased the drug enrichment within biofilms. By loading quorum sensing inhibitors, nanotechnology was also an effective strategy for eradicating bacterial biofilms and recovering the infectious symptoms. Nanobiomaterials could intervene the bacterial metabolism and break the bacterial survival homeostasis by blocking the uptake of nutrients. Moreover, energy-driven micro-nano robotics had shown excellent performance in active delivery and biofilm eradication. Micro-nano robots could penetrate physiological barriers by exogenous or endogenous driving modes such as by biological or chemical methods, ultrasound, and magnetic field, and deliver drugs to the infection sites accurately. Achieving this using conventional drugs was difficult. Overall, the paper described the biological properties and drug-resistant molecular mechanisms of bacterial biofilms, and highlighted therapeutic strategies from different perspectives by nanobiomaterials, such as dispersing bacterial mature biofilms, blocking quorum sensing, inhibiting bacterial metabolism, and energy driving penetration. In addition, we presented the key challenges still faced by nanobiomaterials in combating bacterial biofilm infections. Firstly, the dense structure of EPS caused biofilms spatial heterogeneity and metabolic heterogeneity, which created exacting requirements for the design, construction and preparation process of nanobiomaterials. Secondly, biofilm disruption carried the risk of spread and infection the pathogenic bacteria, which might lead to other infections. Finally, we emphasized the role of nanobiomaterials in the development trends and translational prospects in biofilm treatment.

Surface modification of multifunctional ferrite magnetic nanoparticles and progress in biomedicine / 生物医学工程学杂志

Magnetic ferrite nanoparticles (MFNPs) have great application potential in biomedical fields such as magnetic resonance imaging, targeted drugs, magnetothermal therapy and gene delivery. MFNPs can migrate under the action of a magnetic field and target specific cells or tissues. However, to apply MFNPs to organisms, further modifications on the surface of MFNPs are required. In this paper, the common modification methods of MFNPs are reviewed, their applications in medical fields such as bioimaging, medical detection, and biotherapy are summarized, and the future application directions of MFNPs are further prospected.

Type 2 inflammation in children and advances in its biotherapy / 中华实用儿科临床杂志

Type 2 immunity involves Th2 cells, type 2 innate lymphoid cells(ILC2), and type 2 cytokines such as interleukin (IL)-4, IL-5, and IL-13.Invasion by allergens or harmful substances can cause an excessive type 2 immune response, resulting in type 2 inflammation characterized by the increase in type 2 cytokines, eosinophils, and serum total IgE/specific IgE.Type 2 cytokines are the core of type 2 inflammation, of which production and secretion involve Th2 cells and ILC2.Disruption of barrier function can activate type 2 inflammation and contribute to the vicious cycle.Type 2 inflammation is a common pathophysiological mechanism in many diseases, especially allergic diseases.Type 2 inflammatory disease in children mostly affects the skin, respiratory tract, and gastrointestinal tract.Multiple type 2 inflammatory diseases often co-exist, increasing the disease burden in children.With deepening research on the mechanism of type 2 inflammation, the biotherapy has gained more and more attention.This paper focused on the mechanism, identification and biologics of type 2 inflammation in order to improve clinicians′ understanding of type 2 inflammatory diseases in children.

Progress and prospect of biological treatment for rotator cuff injury repair / 中国修复重建外科杂志

OBJECTIVE@#To review the research progress in biotherapy of rotator cuff injury in recent years, in order to provide help for clinical decision-making of rotator cuff injury treatment.@*METHODS@#The literature related to biotherapy of rotator cuff injury at home and abroad in recent years was widely reviewed, and the mechanism and efficacy of biotherapy for rotator cuff injury were summarized from the aspects of platelet-rich plasma (PRP), growth factors, stem cells, and exosomes.@*RESULTS@#In order to relieve patients' pain, improve upper limb function, and improve quality of life, the treatment of rotator cuff injury experienced an important change from conservative treatment to open surgery to arthroscopic rotator cuff repair. Arthroscopic rotator cuff repair plus a variety of biotherapy methods have become the mainstream of clinical treatment. All kinds of biotherapy methods have ideal mid- and long-term effectiveness in the repair of rotator cuff injury. The biotherapy method to promote the healing of rotator cuff injury is controversial and needs to be further studied.@*CONCLUSION@#All kinds of biotherapy methods show a good effect on the repair of rotator cuff injury. It will be an important research direction to further develop new biotherapy technology and verify its effectiveness.

Interpretation of managing food allergy: Global Allergy and Asthma European Network guideline (2022) / 中华实用儿科临床杂志

Food allergy is one of the most common chronic non-infectious diseases in many countries and regions, which affects 2%-4% of children and adults.Its prevalence is on the rise worldwide.In 2022, the Global Allergy and Asthma European Network (GA 2LEN) proposed recommendations on managing food allergy to people at different age groups.This review aims to interpret the recommendations, clinical practice, precautions, evidence gaps and research priorities of food allergy management based on the GA 2LEN guideline 2022, thus providing reference for clinical management of food allergy.

Hyperadrenocorticism treatment of a dog with high dilution: case report

Hyperadrenocorticism is a common endocrinopathy in dogs, associated to an excessive production or administration of cortisol.Aims:Report the evolution of homeopathic treatment in spontaneous hyperadrenocorticism analyzing with basal and post-acth stimulation cortisol values of a yorkshire male dog with 10 years old, diagnosed from the suppression test with low dose of dexamethasone and stimulation with ACTH, as well as complementary tests.Methodology: Homeopathic treatment was chosen, based on the principle of similitude usingIgnatia amarabecause the patient presents repertorized mental symptoms such as separation anxiety syndrome, docility, annoyances and jealousy and also because it is efficient and less harmful. It was associated to cortisol biotherapy to inhibit cortisol production and adrenal biotherapy to control the response of excess of the cortisol producted by the gland. The exposed information is consented by the tutor.Results:The dog was in convencional treatment with trilostane but didn't response to the therapy, showing 5,41 µg/dL of basal result and 11,8 µg/dL of post-acth result and the symptoms were worst on 12/12/2021, presenting lethargic, panting, more evident alopecia and severe muscle weakness which the patient unable to stand. Therefore, the protocol was recommended for 3 months, included 3 globules ofIgnatia amara30cH orally, every 12 hours, 3 globules of cortisol biotherapy 30 cH and also of adrenal biotherapy 6 cH every 24 hours. On 03/28/2022 the basal and post-acth stimulation results was 3,71 µg/dL e 5,79 µg/dL respectively and the patient was more active, the skin was better and even with difficulty it was movingand having more independence.Conclusion: Homeopathic treatment with high dilution was effective, keeping the indices with the recommended range of post acth between 2.0 and 5.0, confirming an adequate therapeutic monitoring and symptomatic improvement.

Research progress on the role of adipokines in intervertebral disc degeneration / 中国修复重建外科杂志

Objective: To review the research progress of the role and mechanism of adipokines in intervertebral disc degeneration (IVDD) in recent years. Methods: The domestic and foreign literature related to adipokines in the process of IVDD was extensively reviewed. The types and functions of adipokines, the role and mechanism in the process of IVDD, and the application prospects of intervertebral disc biotherapy were reviewed. Results: As a kind of bioactive substance secreted by adipose tissue, adipokine plays an important role in bone and joint diseases, metabolic diseases, and breast cancer. During IVDD, most adipokines can activate multiple signaling pathways by binding to autoreceptors, cause the proliferation and apoptosis of cells and proinflammatory and anti-inflammatory factors parasecretions in the intervertebral disc, and lead to imbalance of intradiscal metabolism and establishment of the initial inflammatory environment, and finally cause the IVDD. Conclusion: Adipokines, as a biologically active substance with metabolic and immunomodulatory functions, play important roles in the occurrence, development, and biological treatment of IVDD.

Photodynamic antimicrobial chemotherapy for repairing a rabbit model of osteomyelitis / 中国组织工程研究

BACKGROUND: Photodynamic antimicrobial chemotherapy holds broad-spectrum antibacterial ability, fast onset of action and non-resistance, which has been extensively applied in the treatment of superficial localized infection. OBJECTIVE: To explore the efficacy of photodynamic antimicrobial chemotherapy for treating osteomyelitis. METHODS: Thirty-six New Zealand white rabbits (provided by Laboratory Animal Center of Academy of Military Medical Sciences) were selected, and a left tibia osteomyelitis model was established. At 28 days after modeling, rabbit models were randomly divided into three groups: Blank group (tibia debridement only), control group (vancomycin/polymethylmethacrylate bone cement was filled into the intramedullary cavity of the infectious tibia after debridement), and experimental group (intramedullary treatment of photodynamic antimicrobial chemotherapy after debridement). The gross observation, imaging examination and bacterial culture were performed at 4, 8, and 12 weeks after surgery. The study was approved by the Ethical Committee of Affiliated Hospital of Logistics College of CAPF, with the approval No. (2015)-0002. RESULTS AND CONCLUSION: (1) Appearance of the skin: Purulent secretion was observed in the blank group, but disappeared in the control and experimental groups where the skin healed well. (2) X-ray examination: With time going, the osteomyelitis aggravated in the blank group. The bone destruction was reduced gradually in the control and experimental groups, and the bone defects healed gradually. There was no significant difference between control and experimental groups at different time points after surgery. (3) Bacterial culture: With time increasing, the bacterial positive rate showed no significant change in the blank group. The bacterial positive rate in the control and experimental groups was on a decline, which showed no significant difference between control and experimental groups at different time points after surgery (P > 0.05). (4) These results indicate that photodynamic antimicrobial chemotherapy is a new treatment for osteomyelitis to effectively control infection, providing experimental basis in clinical practice.

Research progress on periprosthetic joint infection after arthroplasty / 解放军医学杂志

Periprosthetic joint infection (PJI) is a catastrophic complication after artificial joint replacement, and its diagnosis and treatment has always been a great challenge in the field of orthopedics. At present, the treatment strategies for PJI include suppressive antibiotic therapy, debridement antibiotics irrigation of the retained, prosthesis, one-stage revision, two-stage revision, arthrodesis, amputation, and biotherapy, etc. Conventional treatment can not achieve satisfactory results. As a new treatment mode, biotherapy has unique advantages in PJI treatment. This article reviews the risk factors and the source of infection, diagnosis, classification and treatment strategies of PJI, in order to provide valuable reference for clinical treatment of PJI.

Application progress of recombinant human endostatin in the treatment of head and neck cancer / 口腔疾病防治

@#The main treatment of head and neck cancer is comprehensive sequential treatment, but the 5-year overall survival rate is less than 50%. Strategies to further improve the curative effect of head and neck cancer are urgently needed in the clinic. Recombinant human vascular endostatin is an antiangiogenesis drug targeting vascular endothelial cells, which has a certain inhibitory effect on tumors. The treatment of malignant tumors by drugs alone is not significantly better than chemoradiation, but combined with radiotherapy and chemotherapy, it can increase the effect of radiotherapy and chemotherapy without drug resistance by changing the distribution of blood vessels, reducing oxygen and normalizing blood vessels. Head and neck tumor treatment has certain advantages. New tumor treatments are expected. The results of a literature review showed that the mechanism of action of recombinant human endostatin mainly includes regulating the matrix protein inside and outside the endothelial cells to influence neovascularization, acting on receptors related to the surface of endothelial cells, reversing abnormal neovascularization to achieve vascular normalization, inhibiting hypoxia inducible factor to improve the hypoxic status of the tumor area, and regulating the cell cycle to ensure the tumor cells are sensitive to radiation in the sensitive period, and vascular normalization can increase the effect of radiotherapy. This treatment has a good synergistic effect with radiotherapy and chemotherapy of head and neck tumors and has a good effect on advanced head and neck tumors.

Medical treatment and its advances of pancreatic neuroendocrine neoplasms / 中国实用外科杂志

The incidence rate of pancreatic neuroendocrine neoplasms(pNENs)is continuously increasing.Since 20% to64% of pNENs develops metastatic disease when diagnosed and functional pNENs is quite common,medical treatment is of great importance for pNENs.Somatostatin analogues(SSAs)is one of the major medical treatments for pNENs and the new generation of SSAs cannot replace the role of lanreotide autogel and octreotide long-acting repeatable(LAR)for pancreatic neuroendocrine tumor(pNET)with low proliferative index and positive expression of somatostatin receptors(SSTRs).Everolimus and sunitinib are two targeted treatments recommended for pNET.Clinical trials of other targeted treatments are gradually conducted with some of them already reached phase Ⅲ.Chemotherapy is usually applied in pNENs with high growth rate or poorly-differentiated neuroendocrine carcinoma(NEC).Most clinical trials of new chemotherapy scheme are designed for NEC.Peptide receptor radionuclide therapy(PRRT) may have important value in pNET positively expressing SSTRs.Preliminary data of immunotherapy for pNENs did not show very promising result and studies of different medical treatments combinations for pNENs are still under exploration.In general,although medical treatments for pNENs had achieved great advances,there is still a long way to go before many new treatments can be used in pNENs.

Tratamento homeopático em trombocitopenia grave: relato de caso / Homeopathic treatment of severe thrombocytopenia: case report

Descreve-se a evolução do tratamento homeopático de trombocitopenia grave, vinculada ao diagnóstico de erliquiose, em paciente canino fêmea, maltês, de 12 anos de idade, cardiopata, em substituição ao protocolo hospitalar padrão com corticoterapia e transfusão de plaquetas. Em 28/01/2019, a paciente apresentou trombocitopenia de 5 mil/mm³ com petéquias em regiões dorsal e ventral e, ainda, extenso hematoma agudo cervical. O tratamento consistiu na administração sequencial de bioterápico de plaquetas para estimular a produção plaquetária, Phosphorus para conter a hemorragia e 2 doses de Lachesis muta, também para controle hemorrágico. A paciente foi acompanhada e tratada com êxito até 17/04/2019, demostrando eficiência dos medicamentos homeopáticos no tratamento da trombocitopenia grave, sem provocar quaisquer efeitos colaterais. (AU)

We describe a case of homeopathic treatment for severe thrombocytopenia, associated with infection with Ehrlichia canis, corresponding to a 12-year-old Maltese female dog, replacing usual hospital treatment with steroids and platelet transfusion. On 28 January 2019 the patient presented thrombocytopenia­5,000 platelets/mm³­and petechiae on the dorsal and ventral regions, and also an extensive acute cervical hematoma. Treatment consisted of sequential administration of platelet biotherapy to stimulate platelet production and Phosphorus to check bleeding, as well as 2 doses of Lachesis muta also to control bleeding. The patient was followed up until 04/17/2019, the progression demonstrating the efficacy of homeopathic medicines for treatment of severe thrombocytopenia, without causing any side effect. (AU)

Research progress in comprehensive and sequential treatment of tongue squamous cell carcinoma / 口腔疾病防治

@#Tongue squamous cell carcinoma (TSCC) is the most common oral cancer, with early lymph node metastasis and poor prognosis. Surgery is the primary treatment based on sequential therapy for TSCC. The treatment of TSCC has evolved gradually in the past few years and has exhibited a trend of standardization and personalization. Several aspects of TSCC treatment are discussed in this article, such as surgery, radiotherapy, chemotherapy, biotherapy, functional rehabilitation, psychological rehabilitative treatment, prognosis and follow-up systems. This article comments on the types of treatments and research progress for TSCC in China and abroad with the aim of providing a better understanding and references for clinical treatment.

Advances in Medical Treatment for Pancreatic Neuroendocrine Neoplasms / 胃肠病学

Pancreatic neuroendocrine neoplasm (pNEN)is a kind of rare neoplasms with high heterogeneity.Surgery is the first choice to cure local resectable tumor.However,for patients with local advanced tumor or distant metastasis, medical treatment is the main option. Medical treatment mainly encompasses biotherapy, targeted therapy and chemotherapy.Clinicians should make therapeutic option for patients based on the functional status and somatostatin receptor status of the tumor,tumor grade,tumor stage and drug toxicity profile.

Application progress of biotherapy in tumor therapy / 国际生物医学工程杂志

Malignant tumors have become one of the major diseases endangering human health.In recent years,the incidence and mortality of cancer have been increasing.The survival rate is only about 50％ for the cancer patients after common therapists,includes surgery,radiotherapy,and chemotherapy.By facing the severe challenge of choosing treatment method,the more effective cancer treatment has become the focus of clinical researches.Biotherapy is a new tumor therapy method with significant curative effect,and is a new treatment for autoimmune cancer.The application progress of biotherapeutic technologies in cancer therapy were reviewed,including nonspecific immunotherapy,cytokine therapy,adoptive cell therapy,monoclonal antibody therapy and gene therapy.

Preparation and clinical application of esophagus cancer vaccine / 中国综合临床

Objective To explore the anticancer mechanism of esophageal cancer vaccines and clinical effect.Methods Esophagus cancer cells (Eca109) were cultured and the soluble antigen were extracted from the cells.Esophagus cancer vaccine was constructed with the antigen and superantigen SEC.Lymphocytes were isolated from peripheral blood and then stimulated with the vaccine in vitro.Phenotypes of the cells were checked by FCM and killing activity was tested with cytotoxic assays.One hundred and six early esophageal cancer patients were selected after surgery,who were divided into the observation group of 53 cases with esophageal cancer vaccine therapy and 53 cases in the control group with conventional treatment.Then clinical effect was observated and 3 year follow-up survival rate was observed of the of two groups of patients.Results Proliferation of vaccine stimulated lymphocyte group was the strongest,and high peaked at 72 h (A =0.22),and raise CD8+ T cell populations of CTLs.The killing activity of lymphocyte group stimulated by the vaccine against target cells was significantly higher than that of lymphocyte group ((97.36±2.11) ％) vs.(79.27±5.57) ％,F =38.62,P＜0.01).Three years follow up shows that the survival rates of experiment group were 48.27％ (male) and 45.83％ (female) respectively,and control group were 21.43％ (male) and 24.00％ (female),and the difference was significant (x2 =5.06,6.28,P ＜ 0.05).Conclusion The tumor vaccine constructed with esophagus cancer antigen and superantigen SEC can induce PBMC to activate and proliferate into CD8+ CTL with specific cytotoxicity against the cells which the antigen comes from.The vaccine may raise the survival rate of the patients with Esophagus cancer.

Analysis of the effect of clinical application of vaccine for esophageal cancer / 中国肿瘤临床

Objective:To analyze the effects of the clinical applications of vaccine for esophageal cancer to provide treatment ba-sis. Methods:Tumor-soluble antigen was extracted from clinical samples of esophageal cancer tissue. Vaccine for esophageal cancer was prepared using antigen and superantigen staphylococcal enterotoxin C (SEC). One group of patients with esophageal cancer was treated with vaccine after surgery (treatment group). Another group of patients with esophageal cancer was treated using routine meth-ods (control group). After five years, the patients were followed up to analyze survival. Results:After five years of follow up, the sur-vival rates of treatment and control groups were 173/328 (49.71%) and 67/326 (20.55%), respectively. Survival rates significantly dif-fered between the two groups (P<0.05). Furthermore, the survival rates of the female patients were higher than those of the male pa-tients (P<0.05). In the treatment group, the survival rates of patients with low-differentiation cancer were higher than those of patients with high-differentiation cancer (P<0.05). In the control group, the survival rates of patients with low-differentiation cancer were lower than those of patients with high-differentiation cancer (P<0.05). Conclusion:Tumor vaccine prepared with esophageal cancer antigen and superantigen SEC could increase the survival rate of patients with esophageal cancer, particularly female patients and those with low-differentiation cancer.

Strategies of maintenance treatment for patients with advanced non-small-cell lung cancer / 肿瘤研究与临床

Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors,and more than half of newly diagnosed patients with NSCLC are stage Ⅲ B or Ⅳ.Maintenance treatment has been intensively investigated in the field in order to improve time to recurrence and survival time.This paper reviewed recent advances in the maintenance treatment for NSCLC.

In vitro behavior of Mycoplasmagallisepticumlive-type nosode / Comportamento in vitro do bioterápico tipo nosódio vivo mycoplasmagallisepticum / Conducta in vitro de nosodio vivo de mycoplasmagallisepticum

As a step of a doctoral research project, in this study a live-type nosode was prepared from microorganism Mycoplasmagallisepticum strain R (ATCC 93-08/19610) according to Costa model and the rules by Brazilian Homeopathic Pharmacopoeia. Live nosode was tested in vitro to assess safety when used to immunize domestic fowl (Gallus gallus) against infection by this microorganism and to investigate its behavior under laboratory conditions. M. gallisepticum was not shown to grow in fluid (broth) and solid (plate) modified Frey medium with dilutions 11d, 12d, 20d and 30d. Inhibition halos about 2.0 mm were observed around paper disks impregnated with live-type nosode in microorganism-sown Petri dishes, whereas disks impregnated with conventional antibiotic oxytetracycline exhibited 8.0 mm inhibition halos. Protein assessment by Folin-Lowry method showed protein absence in dilutions 12d and 30d and neither microbial DNA traces were found in PCR assay in dilutions 12d, 20d and 30d.

Una etapa de un proyecto de doctorado, en este estudio fue preparado un nosode vivo del microorganismo Mycoplasmagallisepticumcepa R (ATCC 93-08/19610) según el modelo de Costa y las normas de la Farmacopea Homeopática Brasileña. El nosode vivo fue testeado in vitro para determinar su seguridad cuando utilizado para inmunizar aves domésticas (Gallus gallus) contra la infección por este microorganismo e investigar su conducta en condiciones de laboratorio. Fue observado que M. gallisepticum no creció en medio modificado de Frey líquido (caldo) y sólido (placa) con las diluciones 11d, 12d, 20d y 30d. Fueron observados halos de inhibición de aproximadamente 2,0 mm alrededor de discos de papel impregnados con el nosode vivo en placas de Petri sembradas con el microorganismo, mientras fueron observados halos de inhibición de 8,0 mm en los discos impregnados con el antibiótico convencional oxitetraciclina. La medición de proteínas mediante el método de Folin-Lowry evidenció ausencia de proteínas con las diluciones 12d y 30d y ausencia de vestigios de DNA microbiano por PCR con las diluciones 12d, 20d y 30d.

Como parte do experimento de uma tese de doutorado, um bioterápico do tipo nosódio vivo utilizando o microrganismo Mycoplasmagallisepticum estirpe R (ATCC 93-08/19610) foi preparado de acordo com o paradigma desenvolvido por Costa, observando as normas da Farmacopeia Homeopática Brasileira. O nosódio vivo foi testado in vitro com a finalidade de avaliar a segurança de sua utilização para imunizar galinhas domésticas (Gallus gallus) contra a infecção pelo microrganismo M.gallisepticum, e também conhecer seu comportamento sob condições de laboratório. Não houve crescimento de M. gallisepticum no cultivo em meio de Frey modificado líquido (caldo) e sólido (placa) das diluições 11d, 12d, 20d e 30d, enquanto que as diluições 1d e 10d mostraram crescimento do microrganismo. Foram observados halos de inibição de cerca de 2,0 mm em torno de discos de papel impregnados com o nosódio vivo em placas de Petri cultivadas com o microrganismo, enquanto os discos impregnados com o antimicrobiano convencional oxitetraciclina apresentaram halos de inibição de 8,0 mm. A dosagem de proteína pelo método de Folin-Lowry identificou a ausência de proteínas nas diluições 12d e 30d, assim como também não foram encontrados traços de DNA microbiano na prova de PCR para as diluições 12d, 20d e 30d.