eamag/protocols-with-steps huggingface dataset:
TITLE: Western Blotting Protocol
AUTHORS: Sam Li

[DESCRIPTION]
<div class = "text-blocks"></div>

[STEPS]
?. [Sample Preparation:]
Place cells in a microcentrifuge tube and centrifuge to collect the cell pellet.
?. [Sample Preparation:]
Lyse the cell pellet with 100µl of lysis buffer on ice for 30 min (For 1 X 106 cells, lyse with 100µl of lysis buffer).
?. [Sample Preparation:]
Centrifuge at 14,000 rpm (16,000xg) for 10 minutes at 4°C.
?. [Sample Preparation:]
Transfer the supernatant to a new tube and discard the pellet. Remove 20µl of supernatant and mix with 20µl of 2x sample buffer.
?. [Sample Preparation:]
Boil for 5 min. Cool at room temperature for 5 minutes. Microcentrifuge for 5 minutes.
?. [Sample Preparation:]
Load up to 40µl of sample to each well of a 1.5mm thick gel*.*Guidelines for choosing gel percentages are based on protein size to be detected: 4-5% gel, > 200 kD; 7.5% gel, 120-200 kD; 8-10% gel, 40-120 kD; 13% gel, 15-40 kD; 15% gel,
?. [Sample Preparation:]
Set gel running conditions according to the manufacturer’s instructions. Transfer the proteins to a nitrocellulose or PVDF membrane with variable power settings according to the manufacturer’s instructions.
?. [For Amyloid Beta Detection, Boiling Method:]
Immediately after transferring the gel onto the membrane, submerge the membrane in boiling PBS for 5 minutes. After boiling, continue as normal to the membrane blocking step of the protocol.
?. [Membrane Blocking:]
Remove the blotted membrane from the transfer apparatus and immediately place in blocking buffer consisting of 5% nonfat dry milk/TBS-T**.
?. [Membrane Blocking:]
Incubate the blot for 1 hour at room temperature, or overnight at 4°C with agitation.
?. [Antibody Incubation:]
Dilute the primary antibody to the recommended concentration/dilution in 5% nonfat dry milk/TBS-T. Place the membrane in the primary antibody solution and incubate for 2 hours at room temperature, or overnight at 4°C with agitation.
?. [Antibody Incubation:]
Wash three times for 5 minutes each with Wash Buffer (TBS containing 0.1% Tween-20).
?. [Antibody Incubation:]
Incubate the membrane for 30 minutes at room temperature with horseradish peroxidase (HRP)- conjugated secondary antibody, diluted to 1:1000 - 1:5000 in 5% nonfat dry milk/ TBS-T.
?. [Antibody Incubation:]
Wash 4 times for 10 minutes each with TBS containing 0.1% Tween-20 and once for 2 minutes with PBS.
?. [Protein Detection:]
Incubate membrane (protein side up) with 10ml of ECL (enhanced chemiluminescence substrate) for 1-2 minutes. The final volume required is 0.125ml/cm2.
?. [Protein Detection:]
Drain off the excess detection reagent, wrap up the blots, and gently smooth out any air bubbles.
?. [Protein Detection:]
Place the wrapped blots, protein side up, in an X-ray film cassette and expose to x-ray film. Exposures can vary from 5 seconds to 60 minutes.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This medium is used successfully for most cyanobacteria. Vitamin B<sub>12</sub> may be added for those species that require it. Use f/2 vitamin solution.&nbsp;

[GUIDELINES]
<table>
<tbody>
<tr>
<td><strong>STOCK</strong></td>
<td><strong>STOCK SOLUTION</strong></td>
<td><strong>ml/Litre</strong></td>
</tr>
<tr>
<td>NaNO<sub>3</sub></td>
<td>150 g/L</td>
<td>10 ml</td>
</tr>
<tr>
<td>K<sub>2</sub>HPO<sub>4</sub>&middot;3H<sub>2</sub>O or *K<sub>2</sub>HPO<sub>4</sub></td>
<td>40 g/L or *30 g/L</td>
<td>1 ml</td>
</tr>
<tr>
<td>MgSO<sub>4</sub>&middot;7H<sub>2</sub>O</td>
<td>75 g/L</td>
<td>1 ml</td>
</tr>
<tr>
<td>CaCl<sub>2</sub>&middot;2H<sub>2</sub>O</td>
<td>36 g/L</td>
<td>1 ml</td>
</tr>
<tr>
<td>Citric Acid combined with Ferric<br />Ammonium Citrate&nbsp;</td>
<td>6 g/L</td>
<td>1 ml</td>
</tr>
<tr>
<td>EDTA</td>
<td>1 g/L</td>
<td>1 ml</td>
</tr>
<tr>
<td>Na<sub>2</sub>CO<sub>3</sub></td>
<td>20 g/L</td>
<td>1 ml</td>
</tr>
<tr>
<td>Trace Metal Solution</td>
<td>See below</td>
<td>1 ml</td>
</tr>
</tbody>
</table>
<br />Adjust pH to approximately 7.5. (Initial pH is approximately 8.5.) When making solid media, you can add agar directly to medium or make double strength medium and double strength agar solution, then after autoclaving combine the two. OPTION: 0.5 g/L of HEPES buffer can be added to the final medium as a buffer. FeCl<sub>3</sub> and EDTA added in a 1:1 ratio may be substituted.&nbsp;<br /><br /><strong>Trace Metal Solution:&nbsp;</strong><br />
<table>
<tbody>
<tr>
<td><strong>Substance</strong></td>
<td><strong>g/Litre</strong></td>
</tr>
<tr>
<td>H<sub>3</sub>BO<sub>3</sub></td>
<td>2.86 g</td>
</tr>
<tr>
<td>MnCl<sub>2</sub>&middot;4H<sub>2</sub>O</td>
<td>1.81 g</td>
</tr>
<tr>
<td>ZnSO<sub>4</sub>&middot;7H<sub>2</sub>O</td>
<td>0.222 g</td>
</tr>
<tr>
<td>NaMoO<sub>4</sub>&middot;5H<sub>2</sub>O</td>
<td>0.390 g</td>
</tr>
<tr>
<td>CuSO<sub>4</sub>&middot;5H<sub>2</sub>O</td>
<td>0.079 g</td>
</tr>
<tr>
<td>Co(NO<sub>3</sub>)<sub>2</sub>&middot;6H<sub>2</sub>O</td>
<td>0.0494 g</td>
</tr>
</tbody>
</table>
<br />Dissolve each of the above substances separately prior to adding the next on the list.<br /><br />Rippka, R., J. Deruelles, J. Waterbury, M. Herdman and R. Stanier. 1979. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111: 1-61

[STEPS]
TITLE: Data update
AUTHORS: Alise J. Ponsero

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocols aims to details the steps necessary to update previously collected data points for the "Database Justice League" project.</div></div>

[STEPS]
?. [Download data]
Once you have received the link to download your dataset, download the csv file locally.Make sure to save it on your computer in a convenient folder!In this protocol we will perform the data collection on a test dataset:
?. [Download data]
As you can see in this test dataset, some preliminary information have been collected automatically such as the database name,  link and availability in 2016
However some information will need additional efforts to be collected, such as the current availability of the database.
?. [Database availability]
In the dataset, you'll find a column name "current_access". This is the latest link known for the database. In order to access the database, copy and paste this link in your browser.
?. [Database availability]
We know that this ressource was available in 2016. Now, we need to assess if the URL for the database is still working. Click on the URL you identified!Two possibilities: The URL is working or not!
?. [Submit dataset]
Great job! You collected all the data we needed to complete the DB Justice League! You should now have a table filled as follow:Upload the table in your github repo and notify us of the upload!Thank you for your great contribution!
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Protocol for Immunohistochemistry free-floating sections with anti-doublecortin antibody for avian tissue.</p>

[STEPS]
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TITLE: DNA extraction (BOMB)
AUTHORS: Tsu-Chun Hung, Yin-Tse   Huang

[DESCRIPTION]
DNA extraction (BOMB)

[STEPS]
SECTION: Sample Collection
1. Add 200 µL of 1mm beads to 1.5mL eppendorf tube

SECTION: Sample Collection
2. Add200 µL of 0.5mm beads to 1.5mL eppendorf tube

SECTION: Sample Collection
3. Add870 µL Lysis master mix to 1.5mL eppendorf tube

 

SECTION: Sample Collection
4. Collect 10-20 mg of sample to 1.5mL eppendorf tube
SECTION: Sample crush
5. Put 1.5mL eppendorf tube in mixmill for sample crush, at this condition: 30 rpm/s, for 4mins 4 min 
SECTION: Centrifugation
6. Put 1.5mL eppendorf tube in centrifuge for centrifugation, at this condition:17.0 x g, 3 min, 25 °C 
SECTION: DNA purification
7. Add 350 µL of isopropanol to the 1st well of 96 well plate

   
SECTION: DNA purification
7.1. Add 100 µL of magnetic beads (10 mg/ml) to the 1st well of 96 deep well plate 

 


SECTION: DNA purification
7.2. Add 200-300 µL of the sample (lysate) from the 1.5mL centrifuged tube to the 1st well of 96 deep well plate 
 

SECTION: DNA purification
8. Add 400 µL of isopropanol to the 2nd well of 96 deep well plate 
SECTION: DNA purification
9. Add 300 µL of 80% enthanol to the  3rd well of 96 deep well plate 
SECTION: DNA purification
10. Add 300 µL of 80% enthanol to the 4th well of 96 deep well plate 
SECTION: DNA purification
12. Add 100 µL of DEPC-treated water to the 6th well of 96 deep well plate 
SECTION: DNA purification
13. Put the prepared 96 deep well plate in the automated DNA extraction machine and select the BOMB protocol

SECTION: DNA purification
14. After the extraction is done, collect 50-100 µL of the eluted sample as the DNA template for downstream experiments

SECTION: DNA purification
11. Add 300 µL of DDW to the 5th well of 96 deep well plate
TITLE: PCR and analysis
AUTHORS: Enrico Bagnoli, Miratul Muqit

[DESCRIPTION]
This protocol details the PCR analysis.  

[STEPS]
SECTION: PCR setup
1. 
Include at least two housekeeping control
 

SECTION: PCR setup
2. 
 

Prepare a working solution at 10 micromolar (µM) by diluting 1:10 the stock in PCR-grade water.

SECTION: PCR setup
3. In each well of a 384 well-plate,5 µL of samples + master mix have to be added, according to the following volumes:

DNA: 2 µL
MasterMix: 2.5 µL SyberGreen + 0.5 µL forward and reverse primers working stock

Prepare, for each gene used, a master mix, by adding 2.5 µL/well of SYBR green and 0.5 µL/well of primer mix in a clean PCR-grade tube.

SECTION: PCR setup
4. Dilute stock of cDNA. A 1:5 or 1:10 dilution is normally sufficient to detect most genes and to have enough cDNA for all the samples.
SECTION: PCR setup
5. Pipette 2 µL cDNA in each well assigned to the same samples.
SECTION: PCR setup
6. Pipette 3 µL mastermix in each well assigned to the same primer.
SECTION: PCR setup
7. When all plate is completed, seal the plate and quickly centrifuge it to prevent bubble interference.
SECTION: PCR setup
8. Put the plate in a thermal cycler designed for quantitative measurements. 

 

SECTION: PCR analysis
9. The following sections outlined analysis of PCR data using the ΔΔCt method.
SECTION: PCR analysis
10. For each sample and for each gene, calculate the average Ct value of the technical triplicate,, checking that intra-replicate variation is acceptable.
SECTION: PCR analysis
11. For each sample, calculate the geometric mean of the Ct value of the housekeeping genes (Ct HKG)
SECTION: PCR analysis
12. For each sample and for each gene of interest, subtract the average Ct HKG value (from step 11) from the average Ct value calculated at step 10 to obtain the ΔCt GOI for each sample and for each gene of interest
SECTION: PCR analysis
13. For each gene of interest, average the ΔCt GOI for the control samples (untreated) across your experimental replicates to obtain the control ΔCt GOI 
 

SECTION: PCR analysis
14. For each gene of interest, calculate the ΔΔCt GOI by subtracting the control ΔCt GOI from the ΔCt GOI for each sample and for each experimental replicates
SECTION: PCR analysis
15. At this point, you should have, for each gene of interest and for each sample, n ΔΔCt GOI where n is the number of experimental replicates.
 

SECTION: PCR analysis
16. In order to plot the result, use the Fold change for each gene of interest by using the following formula for each data point:

FC=2^(-ΔΔCt GOI)
TITLE: RNA extraction from  Pichia pastoris
AUTHORS: iGEM Dusseldorf

[STEPS]
?. pipeting hole culture  of  Pichia liquid culture into 1.5 mL microcentrifuge tubes
?. centrifugation for 1 min at 10.000 x g
?. discard the supernatant
?. extract RNA with Kit: for e.g. ZymoResearch YeaStar RNA Kit (Cat.#R1002) following the manufacturers manual.
?. messure the concentration
a A260/280 ratio of ~2.0 indicates pure RNA
?. To remove DNA contamination, treated samples with DNase use also a Clean and Concentrator Kit for e.g. the Zymoclean RNA Clean & Concentrator Kit (Cat.# R1013).
?. store RNA directly at -80° C
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol provides an efficient DNA extraction and purification of historical museum hides, which potentially have been chemically tanned.</p>

[BEFORE_START]
<p>Clean</p>

[STEPS]
?. 
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TITLE: shRNA knockdown
AUTHORS: wusj, Nancy C. Hernandez Villegas, schekman

[DESCRIPTION]
This protocol describes a standard procedure to generate a stable cell line for a DNAJC5 knockdown using shRNA. 



[STEPS]
SECTION: shRNA knockdown
2. plKO.1-Hygroplasmids-containing shRNA targeting DNAJC5 (ccggGCAACCTCAGATGACATTAAACTCGAGTTTAATGTCATCTGAGGTTGCTTTTTG) together with pMD2.G and PsPAX2 were transfected into HEK293T cells to produce lentiviral particles for 72 hr. 
SECTION: shRNA knockdown
1. HEK293T cells were plated at a density such that on the day of transfection they are no more than 50% confluent. 

SECTION: shRNA knockdown
3. Lentivirus particles were concentrated with Lenti-X Concentrator follow manufacturer protocol (Takara Bio). 
SECTION: shRNA knockdown
4. SH-SY5Y cells were transduced by lentivirus before differentiation.

SECTION: shRNA knockdown
5. Three days post transduction, cells were selected with 250 μg/ml hygromycin for 10 days. 
SECTION: shRNA knockdown
6. The selected cells were differentiated, and the knockdown was verified with immunoblot.
TITLE: Dishwasher
AUTHORS: Elizabeth Fozo

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Steris Dishwasher protocol</div><div class = "text-block">Steris corporation </div><div class = "text-block">Account manager/rep: LeenaAsplund</div><div class = "text-block"><span>Email: </span><a href="mailto:leena_asplund@steris.com" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;:UNDERLINE;">leena_asplund@steris.com</span></a></div><div class = "text-block">Cell: 603-365-1409</div><div class = "text-block">Fax: 440-350-7077</div><div class = "text-block">Voicemail: 800-989-7575 ext. 20843</div></div>

[STEPS]
?. [Steris Dishwasher protocol]
Open door- bottom left button/door and up arrow graphic
?. [Steris Dishwasher protocol]
Close door- bottom left button/door and down arrow graphic
?. [Steris Dishwasher protocol]
View cycle, Select cycle, and Start cycle are all located on the operation screen
?. [Steris Dishwasher protocol]
Add/delete/change cycles – supervisor login button then enter the password
?. [Steris Dishwasher protocol]
After the cycle starts door will remain locked after the temperature passes 40°C
?. [Standard cycle]
PrewashAlkaline detergent2 rinse cycles with tap water1 final rinse with DI water High heat drying
?. [Heavy cycle]
PrewashAlkaline and Aciddetergent2 rinse cycles with tap water1 final rinse with DI water High heat drying
TITLE: Percutaneous myocardial infarction model in rabbit
AUTHORS: Eline Huethorst, Michael Freeman, Erin Boland, Michael Dunne, Francis Burton, Chris Denning, Rachel Myles, Godfrey Smith

[DESCRIPTION]
This video file contains fluoroscopic recordings of 6 steps of the percutaneous inductions of myocardial infarction in rabbit. 
1. Identification of the aortic structures.
2. Insertion of the guide wire.
3. Guidance of 4F catheter over guide wire. 
4. Identification of the coronary ostium and apical artery. 
5. Engagement of the coronary vessel with a floppy guide wire.
6. Advancement of the micro-catheter and tip over the wire to deploy the tip.

[STEPS]
TITLE: protein extraction from cell pallets
AUTHORS: Jacob   Waldbauer, Lichun Zhang, Jacob   Waldbauer,   Lichun Zhang

[DESCRIPTION]
Use LDS buffer to extract protein from cell palllets

[STEPS]
1. Make 1X LDS buffer. In 40mL of 1X buffer, add Tris HCl 0.666g, Tris base 0.682g, LDS 0.8g, EDTA 0.006g, Glycerol 4g.
2. make 1M DTT stock solution in LC/MS water.
3. Add 200~500uL of 10mM DTT in 1X LDS buffer to the cell pallet tube. 
4. Heat sample up at 95ºC for 20 minutes.
5. Cool down the sample to room temrature. Meanwhile, make a 0.5M iodoacetamide stock solution.
6. Add IAM to 50mM/60mM in the sample, incubate in dark for 30-60minutes.
7. Add DTT to 50mM/60mM. Sampe is ready to continue with FASP.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><strong>Immunochemistry Protocols: Immunopurification ‐ (Small Scale) Using Anti‐Epitope Tag Affinity Matrices:</strong></p>
<p> </p>
<p>Typically, immunoprecipitation of antigen from extracts of transfected mammalian cells requires between 1-5 μL of polyclonal antiserum, 5-100 μL of hybridoma tissue culture medium, or 1-3 μL of ascites. If more antibody is used than is necessary, nonspecific background will increase. The concentration of immunoglobulins in antiserum is approximately 6-8 mg/mL. Supernatant from cultured hybridoma cells typically contains 10-100 μg/mL immunoglobulin, and ascites fluid contains 5-7 mg/mL immunoglobulin.</p>

[GUIDELINES]
<p><strong>*IP Buffer</strong><br />• 50 mm Tris HCl (pH 7.5)<br />• 150 mM NaCl<br />• 0.1% Tween 20 (or 0.1% Nonidet P 40)<br />• 0.5% BSA<br />• 1 mM EDTA (pH 8.0)<br />• 1mM β-mercaptoethanol (see Considerations)<br />• 1mM phenlymethylsulfonyl fluoride (PMSF) or<br />protease inhibitors of choice</p>
<p> </p>
<p><strong>**Wash Buffer</strong><br />• 50 mm Tris HCl (pH 7.5)<br />• 150 mM NaCl<br />• 0.1% Tween 20 (or 0.1% Nonidet P 40)<br />• 1 mM EDTA (pH 8.0)<br />• 1mM phenlymethylsulfonyl fluoride<br />(PMSF) or protease inhibitors of choice</p>
<p> </p>
<p><strong>Considerations:</strong><br />- The ingredient β-mercaptoethanol is not always necessary but is included in the immunopurification buffer to improve the solubility of the proteins in the lysate. At the concentration indicated, it should not reduce antibodies. DTT may be used as a substitute.<br />- The suggested immunopurification buffer is by no means the only one that will work. A wide variety of other common physiologic buffers may be employed instead.<br />- At step 4, if elution is not desired, bound protein may be visualized on a gel by resuspending beads directly in reducing gel loading buffer, boiling for 5 minutes, and loading the supernatants on a gel for analysis. Note that the loading buffer may release a portion of the antibodies from the affinity matrix.</p>

[STEPS]
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TITLE: Midbrain organoid astrocyte seeding 
AUTHORS: gustavo.parfitt

[DESCRIPTION]
Midbrain organoid astrocyte seeding protocol

[STEPS]
1. Refer to the publish protocols to obtain the cell materials required.
https://www.protocols.io/view/midbrain-organoid-differentiation-in-spinner-flask-rm7vzbnr4vx1/v1
https://www.protocols.io/view/astrocyte-extraction-from-brain-organoids-261ge364wl47/v2  
2. From a 90% confluent plate of astrocytes. 
3. Wash with PBS.
4. Add 1 mL  of  TrypLE™ Select Enzyme (1X), no phenol red Thermo Fisher Catalog #12563011.

5. Transfer the cells to 15 ml falcon tubes and add 5 ml of media.
6. 300 rcf, 3 min, 25 °C 

7. Resuspend the cell pellet in 1ml  of  

8. Count the cells using 
 

9. Individually add day 40 midbrain organoids to a Corning  96 well V bottom shape plate.
10. Add 50K astrocyte to the organoids.
11. Add 100 ul of organoid media.
12. After a week transfer the organoids to a 96 well plate coated with   1/100 .
TITLE: Y-Maze Protocol
AUTHORS: Lisa Blackmer-Raynolds, Ian N Krout, Tim Sampson

[DESCRIPTION]
The Y maze is a quick and easy way to evaluate a rodent’s spatial working memory by measuring their ability to effectively explore a Y shape maze. Rodents typically prefer to explore novel arms of the maze, rather than returning to arms they have previously visited. As such, rodents with intact working memory will alternate between all arms of the maze before returning to one they have visited previously.

[STEPS]
SECTION: Methods
1. Bring mice up to the behavior room to acclimate at least 60 min before testing.

SECTION: Methods
2. Clean each maze with 70% ethanol and let dry before beginning each test.

 

SECTION: Methods
3.      
Start Ethovision recording before placing rodents in center of maze (if you have multiple Y mazes, you can run cage mates simultaneously).
SECTION: Methods
4. Leave the room and record for 8 min before returning animals to their home cage.
SECTION: Data Analysis
5. The rodent is considered to have entered an arm when their full body minus their tail is in the arm. 
SECTION: Data Analysis
5.1. Percent alternation is defined as the number of spontaneous alternations/the total possible triads. For example in this maze, if the animal enters the arms in this order: B C A C A B, you would score themas having 2 spontaneous alternations (BCA and CAB) out of 4 possible triads (BCA, CAC, ACA, and CAB) or 50% alternation.
SECTION: Data Analysis
5.2. EthoVision software reports number of alternations and max entries to represent the number of possible triads so percent alternation is simply calculated by number of alternations/max entries. Percent alternation for a healthy mouse should be around 70%.
SECTION: Data Analysis
5.3. Controls: Check that max entries or distance traveled does not correlate with percent alternation. Correlation between these two variables suggests that the amount the rodent is moving may be skewing the percent alternation redouts and is no longer accurately reflecting working memory performance.
TITLE: 0.1xBWT buffer
AUTHORS: Anna Schmidt, Sarah Nagel, Matthias Meyer

[DESCRIPTION]
Protocol for the preparation of 0.1x BWT buffer (Wash buffer II) for automated single-stranded DNA library preparation using the ssDNA2.0 method (Gansauge et al. 2020).

References
Gansauge, M.-T., Aximu-Petri, A., Nagel, S., & Meyer, M. (2020). Manual and automated preparation of single-stranded DNA libraries for the sequencing of DNA from ancient biological remains and other sources of highly degraded DNA. Nature Protocols, 15, 2279-2300. 

[STEPS]
TITLE: Decreased risk of Parkinson’s disease in diabetic patients with thiazolidinediones therapy: An exploratory meta-analysis
AUTHORS: Yueli Zhu, Jiali Pu, Yanxing Chen, Baorong Zhang

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Parkinson’s disease (PD) is a prevalent and complex neurodegenerative disorder with prominent loss of dopaminergic neurons in the substantia nigra. Rest tremor, rigidity, bradykinesia, and postural instability are the four classical motor symptoms. PD is the second most common neurodegenerative disease. The annual incidence is 14/100 000 of the total population in Europe and the Americas, while the rate increased to 160/100 000 in people aged over 65 years. The incidence is similar in Asia. </span><a href="" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">Thiazolidinediones</span></a><span> (TZDs), </span><a href="" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">a class of</span></a><span> peroxisome proliferator-activated receptor-gamma (PPAR-γ) agonists, can improve insulin sensitivity and lower blood glucose level for patients with type 2 diabetes.</span></div><div class = "text-block"><span>Recently, accumulating experiments indicated that TZDs could exert anti-inflammatory and neuroprotective effects in a series of PD animal models. Nevertheless, one randomized controlled trial (RCT) indicated that pioglitazone </span><a href="" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">is unlikely to be able to</span></a><span> slow the disease progression in early PD patients without diabetes. To date, several retrospective observational cohort studies have assessed the association between the TZDs use and the incidence of PD in diabetic patients, but with controversial results.</span></div><div class = "text-block">Thererfore, we performed this meta-analysis to evaluate the efficacy of TZDs in reducing PD risk among diabetic patients. </div></div>

[STEPS]
?.