eamag/protocols-with-steps huggingface dataset:
TITLE: Euplotes crassus transfection through microinjection into the macronucleus
AUTHORS: Rachele Cesaroni

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

[STEPS]
?. Dilute 1:10 Euplotes crassus cultures of two different mating types in artificial sea water (prepare 20 ml culture for each mating type), and feed them with E.coli (3 ml for each mating type). For bacteria preparation see protocol 'Culturing Euplotes crassus to high densities using E.coli as the only food source'.
Recipe for complete seawater (1 L):

36 g Reef Crystals
1 ml Walne's solution
1 ml of 10 μg/ml FeSO40.2 ml of 2 mg/ml thiamine (light sensîtive; store at 4ºC)
Add distilled water up to 1 L
?. Grow cells in 250 ml flat-bottomed flasks at 24°C for 4 days with a 12h light/12h dark cycle, and then mix the same number of cells of both mating types in a 500 ml flat-bottomed flask at room temperature (the optimal cell density for conjugation is ∼1000 cells/ml). Provide no areation in both steps.
?. Isolate single Euplotes crassus cells with a donut shape after 2 days into artificial sea water with 2% BSA, in order to prepare drops for microinjection (ideally one cell each drop).
?. When drops are ready, cover them with a thin layer of Mineral Oil to not let them evaporate.
?. Inject into the macronucleus DNA (3 to 5 μg/μl) using Eppendorf Femtotips I injection needle.
?. Recover each cell individually in 500 μl of artificial sea water plus 0.25 μl of E.coli at 24°C (for bacteria preparation see protocol 'Culturing Euplotes crassus to high densities using E.coli as the only food source').
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div title="Page 1">
<p>Protocol for thymus dissociation (10-week old CD-1 female). </p>
</div>

[GUIDELINES]
<div title="Page 1">
<div>
<div>
<p><strong><em>Bacillus Licheniformis</em> enzyme mix (1 mg/mL enzyme): </strong></p>
<p>492 µL DPBS (No added Ca, Mg)<br /> 0.5 mM EDTA (0.5 μL of 0.5 M EDTA/mL)<br /> 125 U DNAse 1 / mL (2.5 µL)</p>
<p>5 µL of 100 mg/mL enzyme (final conc. 1 mg/mL)</p>
<p>--------------------------------------------<br /> <strong><em>Bacillus Licheniformis</em> enzyme mix (2 mg/mL enzyme): </strong></p>
<p>487 µL DPBS (No added Ca, Mg)</p>
<p>0.5 mM EDTA (0.5 μL of 0.5 M EDTA/mL)<br /> 125 U DNAse 1 / mL (2.5 µL)<br /> 10 µL of 100 mg/mL enzyme (final conc. 2 mg/mL)</p>
<p>--------------------------------------------<br /> +12.5 mg of tissue</p>
</div>
</div>
</div>

[STEPS]
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TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><strong>Bakgrunn</strong></p>
<p>Kvar tredje nye brukar av omsorgstenester blir re-innlagt i sjukehuset, og det er ein samanheng mellom dette, tilhøve ved sjukehus og lengd av opphald. Sjukdom påverkar den helserelaterte livskvaliteten og pasientar si vurdering av helse og funksjon gir viktig informasjon. Pasienterfaringar og re-innlegging av eldre i somatiske sjukehus er to nye kvalitetsindikatorar, indirekte mål på kvalitet innan et område, utgitt av Helsedirektoratet i 2012.</p>
<p> </p>
<p><strong>Hensikt/mål</strong></p>
<p>Formålet med studien er å få kunnskap om eldre pasientars helserelaterte livskvalitet samt erfaringar med sjukehusopphaldet og den første tida etter utskriving.</p>
<p><strong> </strong></p>
<p><strong>Metode</strong></p>
<p>Studien inkluderer 250 menn og kvinner over 65 år med hoftebrot, kols eller hjartesvikt innlagt meir enn 24 timar, og som er i stand til å gje fritt informert samtykke. Deltakarane må vera i stand til å fylle ut spørjeskjema utan hjelp og ikkje ha fast plass i sjukeheim. Metoden er spørjeskjema som kartlegg fysisk og mental helse, angst og depresjon, usikkerheit ved sjukdom, trøttheit og utmatting, subjektive helseplager og erfaringar med sjukehusopphaldet. I tillegg blir det stilt spørsmål knytt til problem etter utskriving, kva type omsorgstenester pasientane mottar, kor dei oppheld seg dei første 30 dagar etter utskriving og re-innlegging i sjukehus i dette tidsrommet. Skjema blir svara på i sjukehuset og ein månad etter utskriving.'' </p>
<p> </p>
<p><strong>Etiske aspekt</strong></p>
<p>Studien blir utført i samsvar med Helsinkideklarasjonen. Det vil bli innhenta samtykke-erklæring frå alle deltakarane før utskriving frå sjukehuset.</p>
<p> </p>
<p><strong>Publisering</strong></p>
<p>Resultat vil bli presentert i internasjonalt tidsskrift med fagfellevurdering.</p>

[STEPS]
TITLE: XP Nutrition Keto Gummies Reviews - Where To Buy It?
AUTHORS: keto-x-reviews  

[DESCRIPTION]
XP Nutrition Keto Gummies Reviews - Where To Buy It?

[STEPS]
SECTION: XP Nutrition Keto Gummies Reviews - Where To Buy It? XP Nutrition Keto Gummies- Reduce Redundant Body Weight.   Dealing with low stamina, poor metabolism position, low energy position, poor digestion, and impunity power and other health issues are some of the health issues which you might face due to rotundity or fat and they&#x27;re common moment and you won&#x27;t overcome from all the health issues fluently and you surely need an effective weight reducing formula which solves different rotundity- related issues and you&#x27;ll gain other benefits also in a healthy way without getting tired. For helping you out we&#x27;ve an advanced result Xp Nutrition Keto Gummies that burns redundant body weight and solves all the colorful weight gaining issues in a healthy way the problem of stress will also get reduced and you&#x27;ll come healthy.   This formula is naturally designed and your impunity and metabolism position will also get bettered so that you noway face any health issues. This formula is chemical-free and provides you with other benefits at the same time and you must read ahead for gaining further information about this product.   About XP Nutrition Keto Gummies   XP Nutrition Keto Gummies is a recently formed fat- burning result that helps you live energetic life by reducing redundant body weight and your stamina and body strength will get bettered. This formula helps you consume healthy food by controlling unwanted hunger and jones. This formula is a chemical-free product and you&#x27;ll only find natural and herbal constituents in it which helps you gain innumerous benefits.   Working on XP Nutrition Keto Gummies   XP Nutrition Keto Gummies simply works for you and improves your health from the inside as this formula enhances the process of ketosis which burns redundant weight from your body and indeed cuts fat from your belly area and improves your energy position noway making you feel lazy and tired. This formula helps in boosting your digestion power which helps your food get digested fluently, impunity power which helps you get healthy from outside and also boosts your metabolism position. It helps you gain advanced stamina, strength, and energy position and controls your hunger, and helps you eat limited and healthy food. It controls your cholesterol, sugar, and blood pressure position and reduces stress situations, and helps you live a healthy and happy life.    Official Website:- https://freaknewswire.com/index.php/2022/05/05/xp-nutrition-keto-gummies-reviews-how-to-use/
1.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol has been optimized to remove washing steps after antibody and nanobeads incubation, resulting in a shorter and more convenient protocol. This procedure is optimized for the isolation of 107 to 2 x 108 cells per tube. If working with fewer cells keep volumes as indicated for 107 cells. For best results, optimize the conditions to your specific cell number and tissue. Prepare fresh MojoSort™ Buffer solution by diluting the 5X concentrate with sterile distilled water.</p>
<p> </p>
<p><strong>Product description and procedure summary</strong>:</p>
<p> </p>
<p>This kit is designed for the sequential positive selection of CD4+ T cells from human peripheral blood mononuclear cells (PBMCs). Human monocytes express both CD14 and CD4. When using only CD4 Nanobeads for positive selection of human CD4 T cells, monocytes could be isolated along with the T cells. If this monocyte fraction does not impact your application, there is no need to address it. However, not including this population may be required. Thus, the first step in this kit is the depletion of CD14+ cells using a combination of biotin anti-human CD14 and Streptavidin Nanobeads. The second step is the positive selection of the CD4 T cells using directly conjugated CD4 Nanobeads. After collection of the targeted cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc.</p>

[GUIDELINES]
<p><strong>Reagents and instruments required:</strong></p>
<p>MojoSort™ Buffer (5X) (Cat. No. 480017)</p>
<p>MojoSort™ Magnet (Cat. No. 480019) or compatible magnetic separation system</p>
<p>Adjustable pipettes</p>
<p>70 μm filters (one per sample)</p>
<p>5 mL (12 x 75 mm) polystyrene tubes</p>
<p>Reagents for sample preparation</p>
<p>Reagents and instruments (Flow cytometer) to determine yield and purity</p>
<p> </p>
<p> </p>
<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/dquecje.jpg" data-src="https://s3.amazonaws.com/pr-journal/dqtecje.jpg" data-ofn="mojosort human cd4 wash.JPG" /></p>
<p>Application notes: To use this product in magnetic separation columns, a titration of the Nanobeads should be performed. Optimal concentration for magnetic separation columns is lot-specific. </p>

[STEPS]
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TITLE: Comprehensive analysis methods for developmental GC exposed zebrafish
AUTHORS: Min-Kyeung Choi, Alex Cook, Helen Eachus, Anna Tochwin, Sara Kuntz, Soojin Ryu

[DESCRIPTION]
This protocol describes methods for the molecular, behavioral, and bioinformatic analyses of GC-exposed zebrafish. All procedures were used for the study; Developmental Glucocorticoid exposure in zebrafish primes expression of genes linked to human psychiatric disorders (Choi et al., 2023)

[STEPS]
TITLE: Preparation of M9 worm buffer
AUTHORS: Alfonso Pérez Escudero, Alid  Al-Asmar, Gabriel Madirolas

[DESCRIPTION]
Preparation of M9 worm buffer, used to maintain Caenorhabditis elegans in liquid for short periods of time. Protocol taken from the WormBook Methods http://www.wormbook.org/chapters/www_strainmaintain/strainmaintain.html

[STEPS]
TITLE: Plasmid construction
AUTHORS: Dan Tudorica

[DESCRIPTION]
Via restriction enzyme digest and ligation

[STEPS]
1. Design PCR primers such that they contain a BglII restriction site upstream of the gene, and SalI downstream of the gene. Design cut sites such that gene will still be in frame post ligation.
2. Acquire purified pmCherryC1 stock, and cleave using BglII and SalI using the standard NEB protocol. Simultaneously cleave PCR product
3. Purify both vector and insert using 1% agarose gel, and compare to uncut controls to make sure that restriction digest went to completion. Gel extract vector and insert.
4. Ligate with T4 ligase at room temperature overnight following standard NEB protocol.
5. Transform into XL10 Gold cells, plate on ampicillin selective medium, grow overnight at 37 C
6. Pick colonies, grow 5 mL overnight culture, and purify plasmid using a commercially available plasmid purification kit.
TITLE: RNA Isolation from Plant Tissue Protocol 4: CTAB-PVP-TRIzol Method
AUTHORS: 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Implemented by: Beijing Genomics Institute</div><div class = "text-block"><span>This protocol is part of a collection of eighteen protocols used to isolate total RNA from plant tissue. (RNA Isolation from Plant Tissue Collection: </span><a href="https://www.protocols.io/view/rna-isolation-from-plant-tissue-439gyr6" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">https://www.protocols.io/view/rna-isolation-from-plant-tissue-439gyr6</span></a><span>) </span></div></div>

[STEPS]
?. Grind tissue to a powder in liquid nitrogen.
?. Add – of ground tissue to  of pre-heated extraction buffer in a 5 ml tube.
200 mg
500 mg
3 ml
?. Vortex the tube until the tissue is mixed with the buffer.
?. Incubate the tube at  for , vortexing briefly () every 2–3 min during the incubation.
65 °C
?. Aliquot the mixture into four 2 ml RNase free tubes,  in each tube.
1 ml
?. Spin the tube at  for  in a centrifuge.
All of the insoluble matter should form a pellet at the bottom of the tube.
?. Pour the supernatant into a new 2 ml tube.
?. Add an equal volume of 24:1 chloroform:isoamyl alcohol to fill the tube.
?. Vortex tubes until the phases mix and appear cloudy.
?. Spin the tubes at  for  in a centrifuge.
?. Transfer the upper aqueous phase to new 2 ml RNase free tubes.
?. Transfer the upper, aqueous phase to new 2 ml RNase free tubes.
?. Spin tubes at  for  in a centrifuge and decant the supernatant, taking care not to lose the pellet.
?. Add  75 % cooled ethanol to the pellet.
1 ml
?. Spin the tube at maximum speed for  in a centrifuge.
?. Open cap and air-dry the pellet.
?. Add  RNase free water to dissolve the pellet.
30 µl
?. Centrifuge at > for .
?. Transfer the upper, aqueous phase to a new 2 ml RNase free tube.
?. Spin tubes at > for  at  in a centrifuge.
4 °C
?. Decant the supernatant taking care not to lose the pellet.
?. Centrifuge at  for  at >.
4 °C
?. Repeat steps 21 and 22 one more time.
?. Open cap and air dry the pellet.
?. Add – RNase free water to dissolve the pellet.
10 µl
30 µl
?. Decant the supernatant carefully.
?. Repeat steps 14 and 15 one more time.
?. Incubate at  for .
20 °C
?. Repeat steps 8 to 10 one more time.
?. Add 1/3 volume of  LiCl to each tube.
?. Mix and let stand at   for – or overnight to precipitate RNA.
4 °C
?. Then add  TRIzol reagent and equal volume of chloroform to TRIzol reagent (Invitrogen).
300 µl
?. Vortex vigorously and store at   for .
20 °C
?. Add 2 volumes of cooled 100% ethanol, 1/10 volume of NaAc and  of glycogen.
2 µl
?. Mix and incubate at  for .
-20 °C
?. Add  75 % ethanol to the pellet.
1 ml
?. Let tube stand at   for .
20 °C
?. Decant the liquid carefully, taking care not to lose the pellet.
?. Briefly centrifuge to collect the residual liquid and remove it with a pipette.
TITLE: Rye Broth Recipe
AUTHORS: Diana Lee

[DESCRIPTION]
This method describes the systematic preparation of a Rye-Based Broth for use in microbiological cultures. The broth consists of rye flour, sucrose, and, optionally, B-sitosterol dissolved in dichloromethane. The method involves the careful mixing of ingredients, filtration, and handling of dichloromethane (optional), followed by autoclaving to sterilize the broth.

[STEPS]
SECTION: Preparation of Rye-Based Broth
1. In a 1.5-liter beaker or jug, combine 60 g of rye flour and 20 g of sucrose with 500 ml of water.
SECTION: Preparation of Rye-Based Broth
2. Mix the contents thoroughly.
SECTION: Preparation of Rye-Based Broth
3. Add 500 ml of boiling water to the mixture. Avoid boiling the solution, as starch may cause viscosity.
SECTION: Preparation of Rye-Based Broth
4. Stir the mixture occasionally for the next 30 minutes to keep the flour in suspension. If using a magnetic stirrer, employ gentle stirring to prevent excessive foaming.
SECTION: Filtration
5. Filter the solution using a fine sieve, avoiding the use of filter paper.
SECTION: Filtration
6. Optionally, for a clear broth, transfer the solution into a 1-liter measuring cylinder and allow it to settle for a few hours to remove rye flour grit.
SECTION: Incorporation of B-sitosterol (Optional)
7. 

Dissolve the B-sitosterol in a small volume of dichloromethane, ensuring complete dissolution. 
 


SECTION: Incorporation of B-sitosterol (Optional)
8. Rapidly add the dissolved B-sitosterol to the rye broth, as dichloromethane evaporates quickly.
SECTION: Incorporation of B-sitosterol (Optional)
9. Employ a magnetic stirrer to gently stir the mixture, ensuring homogeneity
SECTION: Autoclaving
10. Due to the tendency of the rye broth to overflow in the autoclave, fill the Schott bottle no more than half full.
TITLE: Modelling Insulin resistance in vitro
AUTHORS: Laura Mahoney-Sanchez

[DESCRIPTION]
This protocol contains the instruction for modelling insulin resistance in iPSC-derived neurons by depriving the cells of insulin overnight followed by a 30 minutes insulin treatment the following day. 

[STEPS]
1. The media is removed from the cells 
2. Cells are washed 1x with PBS
3. Fresh media is added the cells supplemented with B27 without insulin (Thermofisher A1895601).

4. Cells are left back in the incubator 1080 min  at 37 °C 

5. The following day, insulin was added to the media at either 100 nanomolar (nM) , 250 nanomolar (nM)  or 1 micromolar (µM)  for 30 min
TITLE: Criminal Justice Policy Document Analysis
AUTHORS: Drew Gitomer

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is an adaptation of Yanovitzky & Weber - because we were interested adapting Yanovitzky and Weber's tool for  criminal justice reform.


All of the policymaking process stuff was kept in, but we modified the list of policy actors, and policy focus, and Appendix C.  


We also did our coding in NVIvo so removed Appendix D in the oritinal protocol</div></div>

[STEPS]
TITLE: RNAi feeding 
AUTHORS: Ben Jenkins

[DESCRIPTION]
Daily resuspension and feeding of P. bursaria cultures

[STEPS]
SECTION: Remove supernatant
1. Spin down P. bursaria plate at 800 x g, 10 min, 4 °C

# Ensure that centrifuge is loaded with balance plate.
SECTION: Remove supernatant
2. Transfer P. bursaria plate to OpenTrons OT-2 Liquid Handler.

# Ensure that: P. bursaria plate is in position 7; Fresh waste plate is in position 10; and Pipette rack is refilled with P200 tips in position 1
# Take care when loading the P. bursaria plate into position 7
# Tips can be removed from the outer edges of the Pipette rack beforehand, to save on wastage
SECTION: Remove supernatant
3. Run OT-2 protocol "96-well (1 at at time)" using the OpenTrons App. 

# This removes 150 µL of supernatant
# You may have to manually remove ~75 µL supernatant from Row F once the protocol has finished (multichannel pipette, in the hood)

SECTION: Feeding
5. In a sterile hood, prepare 60 mL NCL containing 120 µL Ampicillin,  240 µL IPTG, and 12 µL β-sitosterol. 

#  Ampcillin (0.1 mg/mL ), IPTG (0.4 millimolar (mM)), and  β-sitosterol (0.0008 mg/mL)
# Conduct the remainder of the experiment under sterile conditions

SECTION: Repeat
4. Return to step 1 and repeat for each plate.
SECTION: Feeding
6. Add 150 µL supplemented NCL to each well. 

# Use a sterile square petri dish and multi-channel pipette

SECTION: Feeding
7. Add 8.3 µL E. coli stock (OD 3) to each well. 

# Use frozen stock plate and multi-channel pipette
# Ensure layout of frozen stock plate matches the P. bursaria plate 
# You can use the same pipette tips for each REPLICATE (so for everything taken from the same row of the stock plate)
# Make sure to pipette stock plate up and down ~20 times (10x in the middle, 10x moving clockwise around the edge) before removing first aliquots from the stock plate
# Pipette up and down gently 3 times before removing aliquot from stock plate, and after adding to P. bursaria plate

SECTION: Feeding
9. Return P. bursaria plate to 23 °C culture room.
SECTION: Repeat
10. Return to step 6 and repeat for each plate.
SECTION: Feeding
8. Pipette P. bursaria plate gently up and down ~20 times to mix cells

# 10x in the middle, 10x moving clockwise around the outside of the well to prevent cells clumping at the edges
# This prevents biofilm formation
TITLE: Cortical neuron differentiation using forced NGN2 expression
AUTHORS: Beatrice Weykopf

[DESCRIPTION]
This is a modified protocol based on (Zhang et al.2013 and Meijer et al., 2019) to generate cryopreserved NGN2 neurons, using hPSCs that carry a doxycline-inducible NGN2 cassette.

The voluming are calculated for 10cm TC dishes and should be adapted accordingly


[STEPS]
SECTION: Overview
1. 
 

SECTION: D0
2. Seed 4x106 single cells onto 1x10 cm dish (coating MG-GFR or with GF 2mg) in iPSC medium supplemented with 10 µM RI in 7ml medium
SECTION: D1
3. Add 8ml KSR with 2µg/ml DOX 
SECTION: D2
4. 9ml 1:1 KSR/N2B w/o B27 2µg/ml DOX and 5µg/ml Puro 

Ensure 24h window between this media change and the previous one
SECTION: D3
5. 10ml N2B  containing 2µg/ml DOX) and 5µg/ml Puro
SECTION: D4
6. 10 ml NBM containing 2µg/ml DOX, 5µg/ml Puro, BDNF 10µg/ml , GDNF 10µg/ml , CNTF 10µg/ml
SECTION: D5
7. 10 ml NBM containing 2µg/ml DOX, 5µg/ml Puro, BDNF 10µg/ml , GDNF 10µg/ml , CNTF 10µg/ml and 10µM DAPT
SECTION: D6 AraC treatment
8. 10 ml NBM 2µg/ml DOX, 5µg/ml Puro, BDNF 10µg/ml, GDNF 10µg/ml, CNTF 10µg/ml, 10 µM DAPT 
add 3 µM AraC varies between cell lines 

! Discard AraC trash according to environmental health guidelines!
SECTION: D7 AraC treatment
9. 10 ml NBM 2µg/ml DOX, 5µg/ml Puro, BDNF 10µg/ml, GDNF 10µg/ml, CNTF 10µg/ml, 10 µM DAPT 
add 3 µM AraC varies between cell lines 

! Discard AraC trash and used medium according to environmental health guidelines!
SECTION: D8
10. 2x wash with 5ml NBM w/o GF to remove AraC completely
add 8-10 ml NBM 2µg/ml DOX, 5µg/ml Puro, BDNF 10µg/ml, GDNF 10µg/ml, CNTF 10µg/ml and
10 µM DAPT 
! Discard AraC trash and used medium according to environmental health guidelines!
SECTION: D9
11. add 8-10 ml NBM 2µg/ml DOX, 5µg/ml Puro, BDNF 10µg/ml, GDNF 10µg/ml, CNTF 10µg/ml and
10 µM DAP
SECTION: D10 Cryopreservation
12. remove medium
SECTION: D10 Cryopreservation
12.1. 1x wash with 4 mLl PBS

SECTION: D10 Cryopreservation
12.2. add 4 mL Accutase supplemented with 10µM RI for 90-120min at37 °C

SECTION: D10 Cryopreservation
12.3. After incubation time; If needed add 200 µL DNAse I

SECTION: D10 Cryopreservation
12.4. Add 3 mL NBM and dissociate using 10 ml serological pipette

SECTION: D10 Cryopreservation
12.5. Pool 3x10cm dishes and collect cells in 1x50 ml falcon after pipetting through a 40µm cell strainer on Ice

!collect all cells in a T175 TC falsk on ice!
SECTION: D10 Cryopreservation
12.6. Rinse dishes with 3 ml NBM
SECTION: D10 Cryopreservation
12.7. Take a sample from the flask to determine cell number
!Always store cells suspension on ice from now on!
SECTION: D10 Cryopreservation
12.8. Aliquot pooled cells into 15 ml falcons and centrifuge at 300g for 5min at 4˚C
SECTION: D10 Cryopreservation
12.9. Freeze cells in 1.5M or 3M aliquots and store at -80˚C overnight and transfer the next day into the liquid nitrogen tank

Freezing medium:
70% KOSRM
20% 1M Trehalose
10% DMSO
TITLE: Investigation of the pain factors of rotator cuff tears: a protocol for a scoping review
AUTHORS: Tomohisa  Yuda, Takashi Saito, Hayato Shigetoh, Takashi  Kitagawa

[DESCRIPTION]
Objective：
The purpose of this scoping review was to investigate the factors associated with rotator cuff tear pain and summarize them into the incidence, exacerbating, and remitting factors.

Introduction：
Rotator cuff tears are one of the most common shoulder disorders. A variety of associated factors have been cited in observational studies, and systematic reviews have examined factors associated with symptomatic rotator cuff tears. However, no reports differentiate pain factors into developmental, exacerbating, and remitting factors.

Inclusion/exclusion criteria：
The study will include patients whose physicians have diagnosed non-traumatic rotator cuff tears using Magnetic Resonance Imaging or ultrasound echo. Outcomes will be a pain, the severity of rotator cuff tear, physical function, and psychological factors. The study design will be observational, with no restrictions on region, race, gender, or language of the original paper.

Methods：
A systematic search of PubMed, Embase, Web of Science, Cumulative Index to Nursing and Allied Health Literature (CINAHL), and Physiotherapy Evidence Database (PEDro) databases using the keywords “rotator cuff tear”, “pain”, and “etiology” will be conducted during May 2022. In the first screening step, two independent reviewers will review all of the titles and abstracts to exclude irrelevant articles. In the second screening step, two independent reviewers will review all of the full texts to exclude irrelevant articles. Outcomes will focus on rotator cuff tear severity, pain, physical function, and psychosocial factors, categorizing factors associated with pain according to each study design and identifying incidence factors, exacerbating factors and remission factors.

[STEPS]