pGM-CSF as an adjuvant in DNA vaccination against SARS-CoV-2.

The emergence of SARS-CoV-2 presents a significant global public health dilemma. Vaccination has long been recognized as the most effective means of preventing the spread of infectious diseases. DNA vaccines have attracted attention due to their safety profile, cost-effectiveness, and ease of production. This study aims to assess the efficacy of plasmid-encoding GM-CSF (pGM-CSF) as an adjuvant to augment the specific humoral and cellular immune response elicited by DNA vaccines based on the receptor-binding domain (RBD) antigen. Compared to the use of plasmid-encoded RBD (pRBD) alone, mice that were immunized with a combination of pRBD and pGM-CSF exhibited significantly elevated levels of RBD-specific antibody titers in serum, BALF, and nasal wash. Furthermore, these mice generated more potent neutralization antibodies against both the wild-type and Omicron pseudovirus, as well as the ancestral virus. In addition, pGM-CSF enhanced pRBD-induced CD4+ and CD8+ T cell responses and promoted central memory T cells storage in the spleen. At the same time, tissue-resident memory T (Trm) cells in the lung also increased significantly, and higher levels of specific responses were maintained 60 days post the final immunization. pGM-CSF may play an adjuvant role by promoting antigen expression, immune cells recruitment and GC B cell responses. In conclusion, pGM-CSF may be an effective adjuvant candidate for the DNA vaccines against SARS-CoV-2.

A novel indolylbenzoquinone compound HL-J6 suppresses biofilm formation and α-toxin secretion in methicillin-resistant Staphylococcus aureus.

Eradication of methicillin-resistant Staphylococcus aureus (MRSA) is challenging due to multi-drug resistance of strains and biofilm formation, the latter of which is an important barrier to the penetration of antibiotics and host defences. As such, there is an urgent need to discover and develop novel agents to fight MRSA-associated infection. In this study, HL-J6, a novel indolylbenzoquinone compound, was shown to inhibit S. aureus strains, with a minimum inhibitory concentration against MRSA252 of 2 µg/mL. Moreover, HL-J6 exhibited potent antibiofilm activity in vitro and was able to kill bacteria in biofilm. In the mouse models of wound infection, HL-J6 treatment reduced the MRSA load significantly and inhibited biofilm formation on the wounds. The potent targets of its antibiofilm activity were explored by real-time reverse transcriptase polymerase chain rection, which indicated that HL-J6 downregulated the transcription levels of sarA, atlAE and icaADBC. Moreover, Western blot results showed that HL-J6 reduced the secretion level of α-toxin, a major virulence factor. These findings indicate that HL-J6 is a promising lead compound for the development of novel drugs against MRSA biofilm infections.

Novel Synthetic Lipopeptides as Potential Mucosal Adjuvants Enhanced SARS-CoV-2 rRBD-Induced Immune Response.

As TLR2 agonists, several lipopeptides had been proved to be candidate vaccine adjuvants. In our previous study, lipopeptides mimicking N-terminal structures of the bacterial lipoproteins were also able to promote antigen-specific immune response. However, the structure-activity relationship of lipopeptides as TLR2 agonists is still unclear. Here, 23 synthetic lipopeptides with the same lipid moiety but different peptide sequences were synthesized, and their TLR2 activities in vitro and mucosal adjuvant effects to OVA were evaluated. LP1-14, LP1-30, LP1-34 and LP2-2 exhibited significantly lower cytotoxicity and stronger TLR2 activity compared with Pam2CSK4, the latter being one of the most potent TLR2 agonists. LP1-34 and LP2-2 assisted OVA to induce more profound specific IgG in sera or sIgA in BALF than Pam2CSK4. Furthermore, the possibility of LP1-34, LP2-2 and Pam2CSK4 as the mucosal adjuvant for the SARS-CoV-2 recombinant RBD (rRBD) was investigated. Intranasally immunized with rRBD plus either the novel lipopeptide or Pam2CSK4 significantly increased the levels of specific serum and respiratory mucosal IgG and IgA, while rRBD alone failed to induce specific immune response due to its low immunogenicity. The novel lipopeptides, especially LP2-2, significantly increased levels of rRBD-induced SARS-CoV-2 neutralizing antibody in sera, BALF and nasal wash. Finally, Support vector machine (SVM) results suggested that charged residues in lipopeptides might be beneficial to the agonist activity, while lipophilic residues might adversely affect the agonistic activity. Figuring out the relationship between peptide sequence in the lipopeptide and its TLR2 activity may lay the foundation for the rational design of novel lipopeptide adjuvant for COVID-19 vaccine.

Advance in Application of Neuroimaging in Plasticity Mechanism after Stroke (review) / 中国康复理论与实践

Neuroimaging technique is a kind of significant means to explore the mechanism of cerebral plasticity after stroke. Diffusion tensor imaging can be used to describe the structure of white matter fiber bundles and evaluate the degree of damage, but it cannot reflect the functional connections between different brain regions. Task-state functional magnetic resonance (fMRI) can detect the activation of corresponding brain regions caused by specific tasks, but the test design is complex and demanding for subjects. Resting-state fMRI can analyze complex brain networks and reflect functional connections in different brain regions, but the method of data analysis is complex. Functional near-infrared spectroscopy (fNIRS) is another non-invasive method to reflect the functional activation of brain regions, in which temporal resolution is better than fMRI, but the spatial resolution is slightly lower. The combination of multiple detection methods may be an important research direction in the future.

Application of Functional Near-infrared Spectroscopy in Brain Area Activation Research: Dominant and Non-dominant Hand under Active Grasp-release Task / 中国康复理论与实践

Objective:To apply functional near-infrared spectroscopy (fNIRS) to analyze brain activity pattern of bilateral sensorimotor cortex (SMC) and premotor cortex (PMC) during complex dominant and non-dominant hand movement in healthy subjects. Methods:From August to December, 2019, 15 right-handed healthy residents were recruited. The block designed grip-release task was used in the subjects, and detected oxyhemoglobin and deoxyhemoglobin concentration with fNIRS to analyze the activation of bilateral SMC, PMC and prefontal cortex in term of activation channels and intensity. Results:For the oxyhemoglobin concentration, the number of activated channels was the same in both hemispheres during right (dominant) hand movement, and the activation of left SMC was stronger (P < 0.05); however, more channels were activated in the right hemisphere during left (non-dominant) hand movement, and the activation of right SMC was stronger (P < 0.05). For the deoxyhemoglobin concentration, more channels were activated in the contralateral hemisphere during either dominant or non-dominant hand movement, and the activation of left SMC, Channel 12 (left PMC) and Channel 26 (right PMC) were stronger during right (dominant) hand movement (P < 0.05). Conclusion:It is feasible to use fNIRS to study the activation of hand movement related brain regions during complex movement of dominant and non-dominant hand, especially with the results of oxyhemoglobin concentration.

Development of Different Methods for Preparing Acinetobacter baumannii Outer Membrane Vesicles Vaccine: Impact of Preparation Method on Protective Efficacy.

Acinetobacter baumannii (A. baumannii) is becoming a common global concern due to the emergence of multi-drug or pan-drug resistant strains. Confronting the issue of antimicrobial resistance by developing vaccines against the resistant pathogen is becoming a common strategy. In this study, different methods for preparing A. baumannii outer membrane vesicles (AbOMVs) vaccines were developed. sOMV (spontaneously released AbOMV) was extracted from the culture supernatant, while SuOMV (sucrose-extracted AbOMV) and nOMV (native AbOMV) were prepared from the bacterial cells. Three AbOMVs exhibited significant differences in yield, particle size, protein composition, and LPS/DNA content. To compare the protective efficacy of the three AbOMVs, groups of mice were immunized either intramuscularly or intranasally with each AbOMV. Vaccination via both routes conferred significant protection against lethal and sub-lethal A. baumannii challenge. Moreover, intranasal vaccination provided more robust protection, which may be attributed to the induction of significant sIgA response in mucosal sites. Among the three AbOMVs, SuOMV elicited the highest level of protective immunity against A. baumannii infection, whether intramuscular or intranasal immunization, which was characterized by the expression of the most profound specific serum IgG or mucosal sIgA. Taken together, the preparation method had a significant effect on the yield, morphology, and composition of AbOMVs, that further influenced the protective effect against A. baumannii infection.

Drug treatment of coronavirus disease 2019 (COVID-19) in China.

Since December 2019, the coronavirus disease 2019 (COVID-19) caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has spread throughout China as well as other countries. More than 8,700,000 confirmed COVID-19 cases have been recorded worldwide so far, with much more cases popping up overseas than those inside. As the initial epicenter in the world, China has been combating the epidemic for a relatively longer period and accumulated valuable experience in prevention and control of COVID-19. This article reviewed the clinical use, mechanism and efficacy of the clinically approved drugs recommended in the Diagnosis and Treatment Protocol for Novel Coronavirus Pneumonia (DTPNCP) released by National Health Commission of P.R.China, and the novel therapeutic agents now undergoing clinical trials approved by China National Medical Products Administration (NMPA) to evaluate experimental treatment for COVID-19. Reviewing the progress in drug development for the treatment against COVID-19 in China may provide insight into the epidemic control in other countries.

[Analysis of traditional Chinese medicine prescriptions based on support vector machine and analytic hierarchy process].

Combined use of drugs is a hot spot in the research of new drugs nowadays, and traditional Chinese medicine (TCM) is a classic practice in the combined use of drugs. In this paper, the compatibility of TCM prescriptions and the related properties of composed herbs were calculated and studied to verify and discuss the feasibility of the results in guiding compatibility. Research Group on New Drug Design, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences had established a structured database of TCM prescriptions by using traditional Chinese medicine inheritance support system (TCMISS V2.0), including 4 012 prescription compatibilities, 2 072 drug components, 381 kinds of TCM diseases, 316 kinds of TCM syndromes and 26 kinds of drug properties. On the basis of the created database above, Support Vector Machine (SVM) was used to analyze the prescription compatibility data and establish a model for predicting feasibility of drug compatibilities. Analytic Hierarchy Process (AHP) and cluster analysis were used to study the influence of drug properties in the rationality of prescription compatibility. The computational results showed that the accuracy in efficacy prediction of two data sets, i.e. prescription-disease and prescription-syndrome, was up to 90% in the linear SVM model. The macro₋averaging and micro₋averaging of the two models were around 0.92, 0.46, respectively. After AHP mapping, most of the incompatible combinations showed significant difference with other drug combinations during the clustering process in the vertical icicle, indicating that the proper machine learning algorithm can be used to lay the foundation for further exploring the combination rules in TCM and establishing more detailed drug-disease and syndrome predicting models, and provide theoretical guidance for the study of the combined use of drugs to a certain degree.

Correlation of gastroesophageal reflux and asthma-like pathophysiological changes / 中华实用儿科临床杂志

Objective To investigate whether gastroesophageal reflux can cause asthma-like pathophysiological changes and its mechanism.Methods Sixty BALB/c mice were randomly divided into 4 groups:group A was gastroesophageal reflux control group,group B was asthma control group,group C was gastroesophageal reflux group,and group D was asthmatic group.The asthmatic models were replicated with ovalbumin(OVA) and aluminum hydroxide,and gastroesophageal reflux models were replicated with hydrochloric acid solution pepsin.After the last inhaling ovalbumin and slow perfusion,the airway pressure was detected,and eosinophil (EOS) and neutrophil granulocyte in bronchial lavage fluid were counted.The flow cytometer was used to determinate IL-4,IFN-γ,and Th1/Th2 ratio changes of spleen cells;Lung tissue and esophagus sections were stained with HE,and pathological changes of lung tissue and esophagus were observed.Results In group C and group D,the airway hyper-responsiveness was significantly increased compared with group A and group B,and the differences were statistically significant (all P ＜ 0.05).In group C and group D,IL-4were significantly increased,while IFN-γand Thl/Th2 ratio were significantly decreased than that in the group A and B group,the differences were statistically significant (all P ＜ 0.05).EOS of group C and group D accounted for a high percentage of total cells and they were significantly higher than that in group A and group B,and the differences were statistically significant (all P ＜ 0.05).Through lung tissue biopsy,in group C and group D,bronchial lumen deformation,infiltration of inflammatory cells around the wall,basement membrane thickening,inflammatory cell infiltration around the wall,peripheral vascular edema,enlarging alveolar cavity,alveolar wall thinning,fracture,part of alveolar fusion into bullace of lung,could be observed.The lung pathological section showed that the endothelial cells in group A and group B were integrated and had no denaturation or necrosis,and there was no inflammation cell infiltrate around.EOS biopsy could be observed in group A and group B of lower esophagus markedly submucosal edema,submucosal inflammatory cell infiltration,and keratin hyper function,visible bacteria group A group B and group D were basic ally normal,with no pathological changes.Conclusions Gastroesophageal reflux can induce Th1/Th2 decreasing,airway hyper-responsiveness and pathophysiological changes similar to asthma.

Effects of diesel exhaust particles inhalation on immediate reaction in asthma rats / 中华儿科杂志

<p><b>OBJECTIVE</b>The role of air pollution on asthma can not be ignored, diesel exhaust particles (DEP) in the air is one of the most important pollutants. This study aimed to investigate the effect and mechanism of DEP inhaled on immediate reaction in the asthma rats.</p><p><b>METHOD</b>Sixty male Wistar rats of "Clean" grade, 6 - 7 week-old, with an average weight of (140 +/- 20) g were used in this study. The rats were randomly divided into 6 groups, 10 in each. Group A was treated with normal saline attack as a negative control, Group B with ovalbumin attack as a positive control. After ovalbumin attack, groups C, D, E, F continued to inhale DEP for 1 week, 2 weeks, 3 weeks and 4 weeks, respectively. The concentration of DEP was 200 microg/ml, the animals were subjected to inhalation of ultrasound nebulized DEP for 30 min per day. One week after all the attacks were concluded, Group A was stimulated with normal saline for 30 min, other groups were stimulated with ovalbumin. Then the airway resistance was determined with multi-channel signal acquisition and processing system and compared. The changes in neutrophils, eosinophils, and other inflammatory cells of BALF and the pathological changes in lung tissue, including epithelial cells loss, the inflammatory cells infiltration around the airway, basement membrane fibrosis, goblet cell hyperplasia etc. were observed. The concentration of IL-5 and gamma-interferon in the lung tissues, and the changes of serum IgE etc. were determined.</p><p><b>RESULT</b>Airway resistance values of group A, B, C, D, E, F after ovalbumin excitation for 30 min were (3.56 +/- 0.21), (7.06 +/- 0.63), (6.46 +/- 0.38), (7.47 +/- 0.33), (8.87 +/- 0.61), (11.00 +/- 0.69) cm H2O/(ml.s). No airway hyperresponsiveness occurred in group A, while Groups B, C, D, E, F had higher airway resistance than group A, group E and F had higher airway resistance than that of group B, the differences were statistically significant. And the airway resistance was different in each group among 0 min, 10 min, 20 min and 30 min (F = 160.646, 148.901, 162.204, 156.186, P < 0.01 for both). The time of DEP inhalation and the airway resistance was positively correlated (r = 0.948, P < 0.01); IgE concentrations of the serum between groups B, C, D, E, F was not significantly different (P > 0.05), but higher than that of group A (F = 2.639, P < 0.01). The infiltrated inflammatory cells included eosinophils and lymphocytes, etc. The percentages of neutrophil(%) were (4.3 +/- 2.0), (9.7 +/- 5.2), (10.3 +/- 5.6), (13.0 +/- 5.2), (42.6 +/- 18.3), (55.3 +/- 6.9). The groups E and F had higher percentage than Group A and Group B (F = 114.226, P < 0.01). The percentages of eosinophils(%) were 0, (11.9 +/- 3.8), (15.8 +/- 6.3), (13.0 +/- 4.9), (21.1 +/- 5.6), (27.1 +/- 4.8). The difference between Groups B, C, D, E, F and Group A was statistically significant. There was significant difference between groups C, D, E, F and group B (F = 46.462, P < 0.05); Lung tissue biopsy in group A showed that the epithelial cells were intact, no inflammatory cells infiltrations were found around the airways, instead, mainly ciliated columnar epithelial cells and only a small number of goblet cells were seen without basement membrane fibrosis. With the inhalation of DEP, the epithelial cells showed gradual necrosis, disruption and loss, goblet cells showed hyperplasia, and infiltrations with inflammatory cells were seen around the airway. In the lung tissue, concentrations of IL-5 in group B, C, and E were (12.8 +/- 2.8), (17.1 +/- 5.2), (18.6 +/- 4.2) pg/mg, the difference between groups C, E and group B was statistically significant (F = 4.236, P < 0.01), the difference in gamma-interferon concentration among all groups was not statistically significance (F = 1.185, P > 0.05).</p><p><b>CONCLUSION</b>DEP inhalation increased the airway responsiveness of asthma rats in immediate reaction, promoted the lung epithelial cell loss, inflammatory cell infiltration, basement membrane fibrosis and goblet cell hyperplasia.</p>

Allergic Pneumonia Animal Model Manufacture / 实用儿科临床杂志

Objective To investigate whether the spores from mushroom antigen can cause the allergic pneumonia and manufacture allergic animal model in the C57BL/_6 mouse.Methods Aged 6 weeks old,weight 25-30 g C57BL/_6 mice were collected.In the mouse tail injection compound of spore antigen and the Freud′s adjuvant.Then pours into through the trachea the antigen once a week.The mice were divided into 4 groups.Group A was the normal mouse,Group B was given Freud′s adjuvant(the same method)to determine whether there was affect to the mouse.Group C and D were injected spore antigen 2 and 4 times.When the antigen sensitization finished 1 week later group C and D were completely divided 2 groups,among them one group was inhalation 1.5% spore antigen and induce the acute response.Six hours later the bronchoalveolar lavage fluid(BALF) was collected to observe cell change,and excise the lung tissue to manufacture the pathology specimen,another group had not been induced the acute response and collect the BALF and to exsise the lung tissue directly.Group B were inhalted saline later to collect the BALF and the lung tissue.In the mouse blood serun,enzyme linked immunosorbent assay(ELISA) was used to mensurate antigen specific IgG.Results In group C and D,antigen specific IgG significantly inhanced than that in group A and B(all P