This work was supported by the National Natural Science Foundation of China (No. 81973919), the National Natural Science Foundation of Guangdong Province (No. 2019A1515011299), and the Innovation and Strong School project of the Department of Education of Guangdong Province (No. 2016KTSCX015).

1.&#160;Protocol registration
<ol>
	<li>Register the systemic review. 
	NOTE: PROSPERO (Table of Materials) was built by National Institute for Health Research for the registration of systematic review. This systemic review protocol has been registered under the number CRD42022301182.</li>
</ol>
2. Software installation
<ol>
	<li>EndNote software installation
	<ol>
		<li>Purchase EndNote software and download the exe&#160;file as an installation program&#160;from EndNote&#39;s official website (Table of Materials).</li>
		<li>Decompress the ZIP file. Double-click to start the installer.</li>
	</ol>
	</li>
	<li>Stata software installation
	<ol>
		<li>Purchase stata software from the Stata official website (Table of Materials).</li>
		<li>Double-click the Stata application program to start the installer.</li>
	</ol>
	</li>
	<li>TSA software installation
	<ol>
		<li>Go to the website of The Copenhagen Trial Unit (Table of Materials) for registration and downloading a ZIP file.</li>
		<li>Double-click the TSA executable jar file for installation.</li>
	</ol>
	</li>
</ol>
3. Database retrieval
<ol>
	<li>Construct retrieval strategy in the following databases: PubMed, Cochrane Library, Embase, Chinese Biomedical Literature Service System (SinoMed), China National Knowledge Infrastructure (CNKI), Chinese Scientific Journals Database (VIP), and WanFang database (Table of Materials).</li>
	<li>Retrieval time: From the inception of the database to July 2023. Take PubMed as an example:
	<ol>
		<li>Use PubMed advanced search builder (Supplementary Figure 1A).</li>
		<li>Combine the search terms as follows: Angong Niuhuang OR Angong pill OR Bezoar pill) AND (Viral encephalitis.</li>
		<li>Download all the retrieved literature: Send To &#62; Citation Manager &#62; Create File (Supplementary Figure 1B).</li>
	</ol>
	</li>
</ol>
4. Inclusion and exclusion criteria
<ol>
	<li>Inclusion criteria
	<ol>
		<li>Study design: Include randomized controlled trials (RCTs).</li>
		<li>Interventions: Include the comparison between Angong Niuhuang Pill (ANP) plus conventional therapy versus conventional therapy alone. 
		NOTE: Conventional therapies involved antibiotics, antiviral drugs, intracranial decompression, antipyretics, and maintaining water-electrolyte balance, etc.</li>
		<li>Patients: Include patients with viral encephalitis based on the diagnostic criteria.
		<ol>
			<li>Altered mental status: Include patients with no other cause found for altered consciousness, drowsiness, or personality changes lasting &#8805;24 h.</li>
			<li>Ensure at least 3 of the following associated manifestations are present: (a) Body temperature higher than 38 &#176;C within 72 h before or after onset; (b) Generalized or partial seizures that cannot be entirely attributed to pre-existing epilepsy; (c) New symptoms of neurological defects; (d) White blood cell count &#8805; 5/mm2 in cerebrospinal fluid; (e) Neuroimaging showed abnormal brain parenchyma; (f) EEG abnormalities consistent with encephalitis that cannot be attributed to other causes.</li>
		</ol>
		</li>
		<li>Outcomes: Include outcomes such as total effective rate, time of defervescence, time of consciousness recovery, time of headache disappearance, and time of tic disappearance.</li>
	</ol>
	</li>
	<li>Exclusion criteria
	<ol>
		<li>Complete the literature retrieval and download the full texts. Read the full texts and exclude studies that meet the exclusion criteria. Record the reasons for each excluded study.
		<ol>
			<li>Additional drug: Exclude studies in which, besides ANP, the treatment group uses additional drugs different from the control group.</li>
			<li>Data missing: Exclude literature with incomplete data.</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>
5. Literature management and screening
<ol>
	<li>Importing literature and removing the duplicates
	<ol>
		<li>Open Endnote and create a new group by clicking My Group &#62; Create Group&#160;(Supplementary Figure 2A).</li>
		<li>Import the retrieved literature by clicking on File &#62; Import &#62; Options &#62; Import Options &#62; EndNote Import &#62; Import (Supplementary Figure 2B).</li>
		<li>Remove the duplicates by clicking Reference &#62; Find Duplicates &#62; Keep this Record (Supplementary Figure 2C).</li>
	</ol>
	</li>
	<li>Screening literature and data extraction
	<ol>
		<li>Create an Excel extraction spreadsheet.</li>
		<li>According to the inclusion and exclusion criteria, have two authors screen the included literature and extract data of included articles independently as follows: 1) first author; 2) publication year; 3) baseline data of a patient with VE (age, gender, ANP usage, duration); 3) outcome measures. 
		NOTE: Record the reasons for each eliminated study.</li>
		<li>Data types: Extract data from dichotomous variables (effective rate), which used sample size and composition ratio, and continuous variables (time of defervescence, time of consciousness recovery, time of headache disappearance, time of tic disappearance), which used mean and standard deviation.</li>
		<li>Create a Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) flow chart, visualizing the number of studies processed at each step and the reasons for removing them.</li>
	</ol>
	</li>
</ol>
6. Quality assessment
<ol>
	<li>Use the Cochrane risk of bias tool to evaluate the methodology. Include the terms of bias assessment: random sequence generation, allocation concealment, blinding of participants and personnel, incomplete outcome data, selective reporting, blinding of outcome assessment, and other biases. 
	NOTE: The third party would intervene to resolve discrepancies by discussions.</li>
</ol>
7. Data synthesis and analysis
<ol>
	<li>Constructing Forest plots using STATA software
	<ol>
		<li>For continuous data, follow steps 7.1.2-7.1.4.</li>
		<li>For data entry, click User &#62; Meta-Analysis of Binary and Continuous &#62; Main &#62; Continuous. Set the sample size, mean, and standard deviation of the treatment group as n1, mean1, and sd1, and set the sample size, mean, and standard deviation of the control group as n2, mean2, and sd2 (Supplementary Figure 3A).</li>
		<li>Effect model: Appy the effect model (fixed/random) based on I2 statistics among the reported trials. I2&#160;&#62; 50% indicates a high heterogeneity; choose the Random model. Click User &#62; Meta-Analysis of Binary and Continuous &#62; Continuous &#62; Random I-V Heterogeneity. Use Fixed Inverse Variance if I2&#160;&#8804; 50%.</li>
		<li>Calculate the outcomes of the same unit as weighted mean difference (WMD); under Statistics, choose noStandard. For different units of outcomes, use the standardized mean difference (SMD); unser Statistics, choose Cohen (Supplementary Figure 3B).</li>
		<li>For dichotomous data, follow steps steps 7.1.6-7.1.7.</li>
		<li>For data entry, use Risk ratios (RR). Click on User &#62; Meta-Analysis of Binary and Continuous &#62; Main &#62; Count. Set the effective number and non-effective number of the treatment group as E1 and NE1, and for the control group as E2 and NE2 (Supplementary Figure 3C).</li>
		<li>Effect model: Apply the effect model (fixed/random) based on I2 statistic among the reported trials. I2&#160;&#62; 50% indicates a high heterogeneity; choose the Random model. Click User &#62; Meta-Analysis of Binary and Continuous &#62; Binary &#62; Fixed. Use Fixed Inverse Variance if I2&#160;&#8804; 50%. 
		NOTE: With small data and few studies, the M-H test is more robust. But M-H can only process binary variables. I-V can be adopted for both continuous and binary variables.</li>
		<li>Performing sensitivity analysis using STATA software
		<ol>
			<li>If I2&#160;&#62; 50% in heterogeneity test, perform sensitive analysis to identify the possible cause of heterogeneity. Click on User &#62; Meta-Analysis &#62; influence analysis, metan-based (metainf) &#62; Continuous (Supplementary Figure 3D).</li>
		</ol>
		</li>
		<li>Checking publication bias using STATA software
		<ol>
			<li>For continuous data, select Egger&#39;s linear regression test to assess the publication bias of continuous variables. Use the command db metabias. Click Main, choose _ES _seES, and select Egger (Supplementary Figure 3E).</li>
			<li>For binary data, use RR or OR as the effect indicators of binary variable Harbord&#39;s weighted linear regression test. Use the command metabias e1 ne1 e2 ne2, or harbord.</li>
		</ol>
		</li>
		<li>Trimming and filling procedure using STATA software
		<ol>
			<li>Visualized by a funnel plot, adopt the trim and fill procedure to test the robustness of the effect size estimate to assess publication bias. Use the command db metatrim.</li>
			<li>Click Main and choose _es_sees. Then, click Linear &#62; Fixed &#62; Funnel (Supplementary Figure 3F). 
			NOTE: Binary variables should make a logarithmic transformation to generate a new dataset for metatrim, such as gen logES = log(_ES).</li>
		</ol>
		</li>
		<li>Trial sequential analysis
		<ol>
			<li>Build a new meta-analysis. Click on File &#62; New Meta-Analysis. Choose Data Type, import Name, Group1, and Group2 and choose the Outcome Type (Supplementary Figure 4A). 
			NOTE: For dichotomous data, negative outcome measures such as death, cancer incidence; positive outcome measures such as survival, viral clearance, or smoking cessation. For continuous variables, a negative mean effect increase represents a negative event (e. g., increase in depression score), and a positive mean increase in mean effect is a positive event (e.g., platelet count).</li>
			<li>Set effect measure and model according to Forest plot results. Use the Constant continuous correction method and Value = 0.5 (Supplementary Figure 4B).</li>
			<li>Click on Trial, fill in the Study, Year, and Effect and Total number of Intervention and Control groups. Then, click Add Trial (Supplementary Figure 4C). 
			NOTE: Supplementary Figure 4D shows the data entry tab that is used to distinguish between continuous and binary variables.</li>
			<li>Click Tsa &#62; Alpha-Spending Boundary. Set Boundary Type as Two-Sided, the Type <img alt="Equation 11" src="/files/ftp_upload/65969/65969eq11.jpg" /> Error rate &#945; = 0. 05 and Power = 0.80.&#160;Click Perform Calculation, then Graph to see the figure.</li>
			<li>For the meta-analysis of dichotomous outcome, fill the expected event rate and intervention effect, including Relative Risk Reduction (RRR) and Incidence in the control group. According to precious forest plot of total effective rate, calculate RRR (Supplementary Figure 4E).</li>
			<li>For continuous data, click Meta-analysis and choose Mean difference and Randon-effect(BT) according the previos results of forest plot.</li>
			<li>Click on Trial, fill in the Study, Year, Mean response, Standard Deviation and Group size of Intervention and Control groups. Then, click Add Trial.</li>
			<li>Click on Tsa. Select Alpha-Spending Boundary. Then, setting the Boundary Type as Two-Sided, the Type &#8544;Error rate &#945; to 0.05 and Power to 0.8. Choose Empirical in mean difference and variance.Click Perform Calculation, then click Graph to see the figure.</li>
		</ol>
		</li>
	</ol>
	</li>
</ol>

Systematic Review and Meta-Analysis on Viral Encephalitis Treatments

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The authors declare that there are no conflicts of interest.

This meta-analysis provides evidence that the adjunctive ANP can improve the total effective rate by 17% in patients with VE. Adjuvant therapy with ANP can significantly shorten the disappearance time of positive pathological signs and symptoms. Xingnaojing (XNJ) injection extracted from ANP is widely used for abnormal consciousness associated with encephalopathy. A recent meta-analysis verified the adjunctive effect of XNJ for treating VE, contributing to an increase in the number of cure cases and shortening the time o...

Encephalitis is defined as the inflammation of the brain parenchyma with high mortality, characterized by neurological pathological signs, fever, abnormal mental status, and altered electroencephalographic (EEG) and neuroimaging1,2. The incidence of encephalitis ranges between 7 and 15 cases per 100,000 people3,4,5. The cause of approximately half of encephalitis cases is unknown, and viral encephalitis (VE) accounts for about 20% to 50% of diagnosed cases5,6. Common viruses include human enterovirus, human herpesvirus, herpes simplex virus, human immunodeficiency virus, and cytomegalovirus7. Patients with VE require intensive monitoring and supportive measures to maintain vital signs and alleviate cerebral inflammation8. Acyclovir is an empiric treatment for all cases of VE9. Evidence for the use of glucocorticoids, immunoglobulins, and interferon alfa are inconsistent8. Angong Niuhuang Pill (ANP), one of the well-known orifice-opening medicines, is widely used in mental and conscious abnormalities due to critical illness. China Medical Education Association recommended ANP as an A-level strength of recommendation and II-level quality of evidence (evidence from non-randomized studies or randomized controlled trials)10.
A systematic review (SR) is a comprehensive collection and rigorous evaluation of all clinical studies that meet the inclusion criteria. Meta-analysis is a methodology for quantitative analysis in SR. Distinguishing from the methods of basic meta-analysis, a trim and fill procedure was adopted to test the robustness of the effect size estimate to the assessment of publication bias11. A small number of studies were removed until a new effect size made the plot symmetrical. Additional studies were added back to the plot based on imputed mirror-image studies. If the difference of the recalculated pooled estimate (PE) before and after the trim and fill procedure was less than 0.05, the effect size estimate of publication bias was robust. TSA 0.9.5.10 software was used to reduce the risk of false-positive results and calculate the required information size (RIS), which was beneficial to make our findings more robust. Given that no systematic review of the data on the improvement of symptoms, signs, biochemical indicators, and adverse reactions of ANP in the treatment of VE, we therefore performed this meta-analysis of the available randomized controlled trials.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>PROSPERO</td>
			<td>National Institute for Health and Care Research</td>
			<td>https://www.crd.york.ac.uk/PROSPERO/</td>
			<td></td>
		</tr>
		<tr>
			<td>STATA</td>
			<td>&#160;StataCorp</td>
			<td>https://www.stata.com/</td>
			<td></td>
		</tr>
		<tr>
			<td>TSA</td>
			<td>Copenhagen Trial Unit, Centre for Clinical Intervention Research</td>
			<td>https://ctu.dk/tsa/downloads/</td>
			<td></td>
		</tr>
		<tr>
			<td>PubMed</td>
			<td>National Library of Medicine</td>
			<td>https://pubmed.ncbi.nlm.nih.gov/</td>
			<td></td>
		</tr>
		<tr>
			<td>Cochrane Library</td>
			<td>Cochrane Library</td>
			<td>https://www.cochranelibrary.com/</td>
			<td></td>
		</tr>
		<tr>
			<td>Embase</td>
			<td>Elsevier</td>
			<td>https://www.embase.com/search/quick</td>
			<td></td>
		</tr>
		<tr>
			<td>SinoMed</td>
			<td>SinoMed</td>
			<td>http://www.sinomed.ac.cn/index.jsp</td>
			<td></td>
		</tr>
		<tr>
			<td>CNKI</td>
			<td>China National Knowledge Infrastructure</td>
			<td>https://www.cnki.net/</td>
			<td></td>
		</tr>
		<tr>
			<td>VIP</td>
			<td>Chongqing VIP Information Co., Ltd.</td>
			<td>http://www.cqvip.com/</td>
			<td></td>
		</tr>
		<tr>
			<td>WanFang database</td>
			<td>Wanfang Data Knowledge Service Platform</td>
			<td>https://www.wanfangdata.com.cn/index.html</td>
			<td></td>
		</tr>
	</tbody>
</table>

the adjuvant efficacy of angong niuhuang pill in the treatment of viral encephalitis: a meta-analysis of randomized controlled trials

This meta-analysis aims to evaluate the efficacy of Angong Niuhuang Pill (ANP) as an adjuvant therapy in the treatment of viral encephalitis. Seven databases (PubMed, Cochrane Library, Embase, SinoMed, CNKI, VIP and WanFang) were included for literature retrieval from inception to July 2023. Randomized controlled trials comparing ANP plus conventional therapy with conventional therapy alone were eligible. Pooled effect sizes and 95% confidence intervals (CIs) were calculated for evaluating efficacy and safety. Sensitivity analysis and publication bias assessments were performed for analyzing the inconclusiveness of findings. 13 studies involving 1045 cases were included for meta-analysis. Adjuvant treatment with ANP increased the probability of the total effective rate by 17% compared with conventional treatment (Risk ratios (RR) = 1.17, 95%CI [1.08, 1.27]). The disappearance time of clinical syndromes and signs was significantly decreased after adjuvant treatment with ANP, including the time of defervescence (weighted mean difference (WMD) = -1.59, 95%CI [-2.09, -1.09]), the time of consciousness recovery (WMD = -1.79, 95%CI [-2.06, -1.51]), the time of headache disappearance (WMD = -1.51, 95%CI [-1.93, -1.08]), the time of tic disappearance (WMD = -1.88, 95%CI [-2.39, -1.36]). The adjuvant efficacy of ANP for treating viral encephalitis (VE) appears to improve the total effective rate and shorten the disappearance time of clinical syndromes. More high-quality randomized controlled trials (RCTs) are needed to support our findings.

Search results and study characteristics 
Systematic retrieval yielded 122&#160;studies and manual searching added 9 relevant literatures. After removing 49&#160;duplicates, 82&#160;studies remained to be screened by abstract and full-text. Ultimately, 13 studies12,13,14,15,16,17,<sup class="x...

Read this Scientific Article Protocol about Author Spotlight: Evaluating the Adjuvant Efficacy and Safety of Angong Niuhuang Pill in Viral Encephalitis Treatment at JoVE.com

Author Spotlight: Evaluating the Adjuvant Efficacy and Safety of Angong Niuhuang Pill in Viral Encephalitis Treatment

Here, we present a protocol to analyze the effectiveness and safety of Angong Niuhuang Pill in the treatment of viral encephalitis.

The Adjuvant Efficacy of Angong Niuhuang Pill in the Treatment of Viral Encephalitis: A Meta-Analysis of Randomized Controlled Trials

This meta-analysis aims to evaluate the efficacy of Angong Niuhuang Pill (ANP) as an adjuvant therapy in the treatment of viral encephalitis. Seven ...

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387 Views.  Guangzhou University of Chinese Medicine. Here, we present a protocol to analyze the effectiveness and safety of Angong Niuhuang Pill in the treatment of viral encephalitis.

Video: Author Spotlight: Evaluating the Adjuvant Efficacy and Safety of Angong Niuhuang Pill in Viral Encephalitis Treatment

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these diseases. Chronic retinal diseases such as retinal oedema or neovascularization usually require multiple intraocular injections to effectively treat the condition. However, the risks associated with these injections increase with repeated delivery of the drug. Therefore, alternative delivery methods need to be established in order to minimize the risks of reinjection. Several other investigations have developed methods to deliver drugs over extended time, through materials capable of releasing chemicals slowly into the eye. In this investigation, we outline the use of Elvax 40W, a copolymer resin, to act as a vehicle for drug delivery to the adult rat retina. The resin is made and loaded with the drug. The drug-resin complex is then implanted into the vitreous cavity, where it will slowly release the drug over time. This method was tested using 2-amino-4-phosphonobutyrate (APB), a glutamate analogue that blocks the light response of the retina. It was demonstrated that the APB was slowly released from the resin, and was able to block the retinal response by 7 days after implantation. This indicates that slow-release drug delivery using this copolymer resin is effective for treating the retina, and could be used therapeutically with further testing.

This paper details how Elvax 40W can be used as a slow-release method for drug delivery to the adult rat retina. The protocol for preparing, loading, and delivering the drug-resin complex to the eye is described.

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Testing the Efficacy of Pharmacological Agents in a Pericardial Target Delivery Model in the Swine

To date, many pharmacological agents used to treat or prevent arrhythmias in open-heart cases create undesired systemic side effects. For example, antiarrhythmic drugs administered intravenously can produce drops in systemic pressure in the already compromised cardiac patient. While performing open-heart procedures, surgeons will often either create a small port or form a pericardial cradle to create suitable fields for operation. This access yields opportunities for target pharmacological delivery (antiarrhythmic or ischemic preconditioning agents) directly to the myocardial tissue without undesired side effects.
We have developed a swine model for testing pharmacological agents for target delivery within the pericardial fluid. While fully anesthetized, each animal was instrumented with a Swan-Ganz catheter as well as left and right ventricle pressure catheters, and pacing leads were placed in the right atrial appendage and the right ventricle. A medial sternotomy was then performed and a pericardial access cradle was created; a plunge pacing lead was placed in the left atrial appendage and a bipolar pacing lead was placed in the left ventricle. Utilizing a programmer and a cardiac mapping system, the refractory period of the atrioventricular node (AVN), atria and ventricles was determined. In addition, atrial fibrillation (AF) induction was produced utilizing a Grass stimulator and time in AF was observed. These measurements were performed prior to treatment, as well as 30 min&#160;and 60 min&#160;after pericardial treatment. Additional time points were added for selected studies. The heart was then cardiopleged and reanimated in a four chamber working mode. Pressure measurements and function were recorded for 1 hr after reanimation. This treatment strategy model allowed us to observe the effects of pharmacological agents that may decrease the incidence of cardiac arrhythmias and/or ischemic damage, during and after open-heart surgery.

We have developed a swine model for the target delivery of pharmacological agents within the pericardial space/fluid. Using this approach, the relative benefits of administered agents on induced atrial fibrillation, relative refractory periods and/or ischemic protection can be investigated.

To date, many pharmacological agents used to treat or prevent arrhythmias in open-heart cases create undesired systemic side effects. For example, ...

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold standard treatment for the high-grade non-muscle invasive bladder cancer (NMIBC). However, since the treatment-related side effects are relevant, newer biological response modifiers with a better benefit/side effects ratio are needed.
The tumour microenvironment can influence both tumour development and therapy efficacy. In order to obtain a good model, it is desirable to implant tumour cells in the organ from which the cancer originates.
In this protocol, we describe a method for establishing a tumour in the bladder cavity of female mice and subsequent delivery of therapeutic agents; the latter are exemplified by our use of Helicobacter pylori neutrophil activating protein (HP-NAP). A preliminary chemical burn of the mucosa, followed by the injection of mouse urothelial carcinoma cell line MB49 via urethral catheterization, enables the cells to attach to the bladder mucosa. After a period, required to allow an initial proliferation of the cells, mice are treated with HP-NAP, administrated again via catheterization. The anti-tumour activity of HP-NAP is evaluated comparing the tumour volume, the extent of necrosis and the degree of vascularization between vehicle- and HP-NAP-treated animals.

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Here the method to establish a syngeneic mouse model of orthotopic bladder tumour to evaluate the anti-tumour efficacy of the bacterial protein HP-NAP is described.

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold ...

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control seizures. However, a significant minority of patients continues to exhibit post-operative seizures, even in those cases in which the suspected source of seizures has been correctly localized and resected. The protocol presented here combines a clinical procedure routinely employed during the pre-operative evaluation of temporal lobe epilepsy (TLE) patients with a novel technique for network analysis. The method allows for the evaluation of the temporal evolution of mesial network parameters. The bilateral insertion of foramen ovale electrodes (FOE) into the ambient cistern simultaneously records electrocortical activity at several mesial areas in the temporal lobe. Furthermore, network methodology applied to the recorded time series tracks the temporal evolution of the mesial networks both interictally and during the seizures. In this way, the presented protocol offers a unique way to visualize and quantify measures that considers the relationships between several mesial areas instead of a single area.

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is often capped with biocompatible materials in order to expedite pulpal wound healing. The ultimate goal is to regenerate reparative dentin, a physical barrier that functions as a "biological seal" and protects the underlying pulp tissue. Although this direct pulp-capping procedure has long been used in dentistry, the underlying molecular mechanism of pulpal wound healing and reparative dentin formation is still poorly understood. To induce reparative dentin, pulp capping has been performed experimentally in large animals, but less so in mice, presumably due to their small sizes and the ensuing technical difficulties. Here, we present a detailed, step-by-step method of performing a pulp-capping procedure in mice, including the preparation of a Class-I-like cavity, the placement of pulp-capping materials, and the restoration procedure using dental composite. Our pulp-capping mouse model will be instrumental in investigating the fundamental molecular mechanisms of pulpal wound healing in the context of reparative dentin in vivo by enabling the use of transgenic or knockout mice that are widely available in the research community.

We describe a step-by-step method of performing direct pulp capping on mice teeth for the evaluation of pulpal wound healing and reparative dentin formation in vivo.

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...

Assessment of the Efficacy of An Osteopathic Treatment in Infants with Biomechanical Impairments to Suckling

Breastfeeding can be challenging for mother-infant dyads experiencing biomechanical suckling difficulties. Although lactation consultants (LCs) all over the world have increased their skills in this field and can provide support to help position the infant at the breast, the impact of their intervention might be limited in the presence of stiff structures in the infants. Here we present a protocol for a randomized controlled trial to assess the efficacy of osteopathic treatment, coupled with lactation consultation, for infants&#39; biomechanical suckling difficulties. It proposes a set-up and a sequence of actions to ensure an optimal context for treatment, as well as a blinding of parents and LCs to the intervention. Data such as the infant&#39;s latch ability measured with the LATCH Assessment Tool, the mother&#39;s nipple pain with a visual analog scale (VAS), and the mother&#39;s perceptions are collected by LCs four times over a 10-day period. Osteopathic lesions are documented by the osteopath, using a standardized assessment grid. Structures of interest are coherent with the anatomical zones involved in latching onto the breast. This protocol also proposes a strategy to document systematically an osteopathic profile of infants with biomechanical suckling difficulties in their first weeks of life. The implementation of this protocol confirms its feasibility for osteopathic assessment and treatment and paves the way for future trials to further explore the efficacy of osteopathic techniques for infants with biomechanical suckling difficulties.

Osteopathy is an emerging field of clinical research. Here we present a protocol to assess the efficacy of an osteopathic intervention coupled with lactation consultation, in infants with biomechanical issues impeding breastfeeding.

Breastfeeding can be challenging for mother-infant dyads experiencing biomechanical suckling difficulties. Although lactation consultants (LCs) all over ...

Large Animal Model for Evaluating the Efficacy of the Gene Therapy in Ischemic Heart

Coronary artery disease is one of the significant causes of mortality and morbidity worldwide. Despite the progression of current therapeutics, a considerable proportion of coronary artery disease patients remain symptomatic. Gene therapy-mediated therapeutic angiogenesis offers a novel therapeutic method for improving myocardial perfusion and relieving symptoms. Gene therapy with different angiogenic factors has been studied in few clinical trials. Due to the novelty of the method, the progress of myocardial gene therapy is a continuous path from bench to bedside. Therefore, large animal models are needed for evaluating the safety and efficacy. The more the large animal model identifies the original disease and the endpoints used in clinics, the more predictable outcomes are from clinical trials. Here, we introduce a large animal model for evaluating the efficacy of the gene therapy in the ischemic porcine heart. We use clinically relevant imaging methods such as ultrasound imaging and 15H2O-PET. For targeting the gene transfers into the desired area, electroanatomical mapping is used. The aim of this method is: (1) to mimic chronic coronary artery disease, (2) to induce therapeutic angiogenesis at hypoxic areas of the heart, and (3) to evaluate the safety and efficacy of the gene therapy by using relevant endpoints.

Myocardial gene therapy for ischemic heart disease holds great promise for future therapeutics. Here, we introduce a large animal model for evaluating the efficacy of gene therapy in the ischemic heart.

Coronary artery disease is one of the significant causes of mortality and morbidity worldwide. Despite the progression of current therapeutics, a ...

Laparoscopy-endoscopy Cooperative Surgery for the Treatment of Gastric Gastrointestinal Stromal Tumors

In view of the shortcomings of endoscopic or laparoscopic surgeries alone in the treatment of gastric gastrointestinal stromal tumors (G-GISTs), this approach makes an innovative improvement in the treatment of G-GISTs that are less than 5 cm in size. Laparoscopy-endoscopy cooperative surgery (LECS) is used to combine endoscopic and laparoscopic surgeries, fully realizing their respective advantages and avoiding their drawbacks. The main steps are as follows. First, gastroscopy and laparoscopy are combined to confirm the location and boundary of the tumor. Tumor resection is carried out laparoscopically, guided by a gastroscope. The specimen is removed orally and the gastric wound closed laparoscopically. Then, gastroscopy and laparoscopy are combined to determine whether there is wound bleeding, if the suture is satisfactory, and if the gastric cavity is deformed.
LECS has natural advantages in the treatment of G-GISTs that are less than 5 cm in size. The accurate estimation of tumor location and boundary greatly improves the complete resection rate of tumors. The risk of tumor rupture is substantially reduced, and the long-term prognosis of patients is significantly improved. The process allows for accurate resection of the tumor, maximum preservation of normal gastric tissue and organ function, and avoids postoperative gastric deformation. The patient's postoperative rehabilitation is greatly accelerated, and oral feeding can resume on the day of the operation. The specimen is taken out through the mouth to avoid the need for an extended abdominal incision. This greatly reduces the patient's postoperative pain and scarring. The method greatly shortens the postoperative hospital stay (i.e., discharge is possible on the day after the operation), increasing the turnover of hospital beds.

Laparoscopy-endoscopy cooperative surgery is appropriate for the treatment of gastric gastrointestinal stromal tumors less than 5 cm in size. It can achieve the respective advantages of endoscopic and laparoscopic surgeries while avoiding their drawbacks.

In view of the shortcomings of endoscopic or laparoscopic surgeries alone in the treatment of gastric gastrointestinal stromal tumors (G-GISTs), this ...

The Adjuvant Efficacy of Angong Niuhuang Pill in the Treatment of Viral Encephalitis: A Meta-Analysis of Randomized Controlled Trials

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The Measurement and Treatment of Suppression in Amblyopia

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Testing the Efficacy of Pharmacological Agents in a Pericardial Target Delivery Model in the Swine

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

Assessment of the Efficacy of An Osteopathic Treatment in Infants with Biomechanical Impairments to Suckling

Large Animal Model for Evaluating the Efficacy of the Gene Therapy in Ischemic Heart

Laparoscopy-endoscopy Cooperative Surgery for the Treatment of Gastric Gastrointestinal Stromal Tumors