Implementation of emergency endovascular aortic repair provides an attractive opportunity in the treatment of complicated acute aortic syndromes involving descending aorta.  The aim of this study was to analyze the effectiveness of thoracic endovascular aortic repair (TEVAR) for the treatment of acute surgical emergencies involving the descending thoracic aorta.  A retrospective review of the medical records of all patients undergoing TEVAR in a single center since 2007 was undertaken. Patients with the aortic disease treated on emergency inclusion criteria were complicated spontaneous acute aortic syndrome (csAAS), traumatic aortic acute injuries (TAIs), and other indications requiring emergent intervention. Technical and clinical success with patient mortality, survival, and reoperation rate was evaluated according to Society for Vascular Surgery reporting standards for thoracic endovascular aortic repair (TEVAR). n = 48) and traumatic aortic acute injuries (TAIs) (31.1%). In addition, in one case aortic iatrogenic dissection (AID) and in 2 other fistulas after the previous stent graft, implantations were diagnosed. All procedures were done through surgically exposed femoral arteries while 2 hybrid procedures required additional approaches. The primary technical success rate was 95.9%, in 3 cases endoleak was reported. The primary clinical success occurred in 94.5%. All patients survived the endovascular interventions, whereas during in-hospital stay one of them died due to multiorgan failure . During the follow-up period, lasting 6 through 164 months (median 67), 11 patients died. Annual, five- and ten-year probability of survival was 86.4 ± 0.04%, 80.0 ± 0.05%, and 76.6 ± 0.06%, respectively. However, the rate of 5-year survivors was significantly higher after TAI (95.2%) than scAAS (63.4%) (p=0.008). Early after the procedure, one individual developed transient paraparesis (1.3%). No other serious stent-graft-related adverse events were noted within the postdischarge follow-up period.  The emergency interventions were necessary in 74 cases (51.0%), including patients with the complicated spontaneous acute aortic syndrome (csAAS) (64.8%;  Descending aortic pathologies requiring emergent interventions can be treated by endovascular techniques with optimal results and low morbidity and mortality in an experienced and dedicated team. Thoracic endovascular aortic repair (TEVAR) initially developed for elective procedures has also become an attractive method of treating acute descending aortic syndromes (AASs) in emergency indications. Conventional open surgery is associated with high perioperative mortality and morbidity. TEVAR application led to a positive decline in the operative mortality and morbidity. The most common forms with poor prognosis and the necessity of emergent interventions are complicated aortic dissection and disruption , 2. CompAcute aortic diseases, particularly those confining also descending part, must be considered as an interdisciplinary entity that should involve specialists in surgery (both cardiac and vascular), interventional radiology, anesthesiology, and cardiology. Such approach through the acceleration of the diagnostic and therapeutic process of critically ill patients with multiorgan trauma directly translates into the results of treatment and final outcomes.This is the first single-center retrospective, observational, cohort study that provides technical and clinical success according to Society for Vascular Surgery reporting standards for thoracic endovascular aortic repair (TEVAR) . MoreoveThe aim of this study was to analyze the effectiveness of thoracic endovascular aortic repair (TEVAR) for the treatment of acute surgical emergencies involving the descending thoracic aorta catastrophes.A retrospective review of the medical records of all patients undergoing TEVAR in a single center since 2007 was undertaken Since 2007, 145 patients with descending aorta pathologies have been treated by TEVAR in the Department of Cardiac Surgery and Transplantology, a center that served as referential TEVAR center for nearly 5 million of inhabitants. Among them, in 74 cases (51%), the emergency TEVAR procedures, defined as emergent interventions (starting before the beginning of the next working day) were performed. Patients with the aortic disease treated on an emergency inclusion criteria were complicated spontaneous acute aortic syndrome (csAAS), traumatic aortic acute injuries (TAIs), and other indications requiring emergent intervention. Patients with elective descending aorta aneurysm (DAA), uncomplicated Type B aortic dissections, and requiring elective TEVAR reinterventions were excluded. The STROBE checklist  is appliAccording to the rules of the Local Bioethical Committee of our university, the Statement of Ethics Approval is not required for retrospective data analysis of patients treated with the use of the standard and accepted methods.From each patient we collected preoperative information, specific data according to the indication of repair, intraoperative data, and postoperative data. Technical and clinical success with patient mortality, survival, and reoperation rate were evaluated according to Society for Vascular Surgery reporting standards for thoracic endovascular aortic repair (TEVAR) . We specThese clinical complications were: infection, postoperative bleeding requiring repeat surgery, postoperative paraplegia, cerebrovascular accident (CVA), acute coronary events, renal failure with a need for dialysis, respiratory failure, and intestinal ischemia. We also analyzed survival at 30 days and 6–164 months follow-up period.The approval in the Department protocol after thoracic stent graft implantation included physical examinations completed by computed tomographic angiography (CTA) at 1, 3, 6, and 12 months after the procedure and later on once a year. Patients who had the hybrid procedures were additionally followed up every 3 months for the first 3 years.Emergent Interventions. Operation before the beginning of the next working day after the decision to operate (EuroSCORE).Critical Preoperative State. ≥1 of the following in the same hospital admission as the operation: ventricular tachycardia or fibrillation or aborted sudden death; cardiac massage; ventilation before arrival to OR; inotropes; IABP or VAD before arrival to OR; acute renal failure, defined as anuria or oliguria <10 mL/hr (EuroSCORE).Post-Implantation Syndrome (PIS). The Velazquez syndrome, called also postimplantation syndrome (PIS), was defined by the presence of leukocytosis  and the occurrence of fever (>38°C–auricular temperature) but without following markers of infection such as increased concentrations of high sensitivity C-reactive protein and procalcitonin when infectious complications were excluded  [l wound) .Primary Technical Success. This is defined on an intent-to-treat basis that begins with the implantation procedure and requires the successful introduction and deployment of the device in the absence of surgical conversion to open repair, death <24 hours, type I or III endoleaks as evidenced by procedural angiography, or graft obstruction [Primary Clinical Success. This can only occur without any of the following: death as a result of treatment or as a result of the original pathology that was treated; type I or III endoleak, infection, or aortic thrombosis; aneurysm expansion  or rupture; conversion to open repair; or failure to arrest the original pathologic process  or causing a new thoracic aortic pathology as a result of the intervention  [ematoma) ;Secondary Clinical Success: This is defined as clinical success obtained initially but temporarily interrupted by a failure that is corrected with the use of an additional, secondary surgical procedure; for example, a type I endoleak develops in an initially excluded aneurysm due to endograft migration at 2 years and is corrected by placement of a new, more proximal endograft. Conversely, clinical failure includes death as a result of treatment or as a result of the original pathology that was treated or a pathology caused by the initial procedure , type I or III endoleak, graft migration, infection, or thrombosis, aneurysm expansion , conversion to open repair, failure to arrest the original pathologic process, or appearance of a new thoracic aortic pathology as a result of intervention [rvention ;All endovascular procedures were performed by the experienced team of two cardiac surgeons and one interventional radiologist, mostly in the vascular intervention room. An exception was two complex procedures carried out in the hybrid room with the assistance of a vascular surgeon. In addition, all but two were treated under general anesthesia. These two patients had to undergo procedures with local anesthesia and sedation but without endotracheal intubation because of diagnosed tracheal injury. Standard antibiotic prophylaxis was used and all patients obligatory received 5,000 IU of heparin. An exception was one patient after trauma with a fracture of the skull base. In all cases, a stent graft was implanted by means of a femoral surgical approach through the right common femoral artery. The left femoral artery was punctured percutaneously by Seldinger's technique for the introduction of a 6F straight catheter with side holes on a pigtail 5F one in the ascending aorta to position the landing zone target. The stent grafts were placed in the thoracic aorta over an Amplatz 0.35 guidewire. DSA  was performed in five to ten series with the use of an ionic contrast medium to confirm the final position and tightness of the prosthesis.In two cases, after CT examination analysis multidisciplinary team decided to perform a two-step procedure in a hybrid room. After carotid-to-carotid anastomoses with FEP Ringed GORE-TEX STRETCH  vascular graft prostheses (8 mm in diameter), eventually the stent grafts were implanted distally to brachiocephalic trunk but covering the orifices of both the left common carotid and left subclavian arteries.n) with percentages (%). The probability of survival was stratified according to the Kaplan–Meier method and if applicable, compared by means of the Cox F test. p value less than 0.05 was considered to indicate differences of statistical significance. The statistical analysis was performed by the use of Statistica 13.3 .The quantitative variables were checked for normality distribution with the use of the Shapiro–Wilk W test. If they met normal distribution they were presented as mean with standard deviation, otherwise as median with range . Categorical variables were expressed as the numbers , the same number with ruptured descending thoracic aortic aneurysms (RTAA) and 2 (2.7%), one with penetrating atherosclerotic ulcers (PAU) and one intramural hematoma (IH). In 23 cases (31.1%) indication for emergent TEVAR was (31.1%) TAI, and among them 14 with traumatic aortic disruption (TAD). In one case aortic iatrogenic dissection (AID) with mesenteric ischemia and in the other 2 subjects aorto-oesophageal fistulas were the other indications for emergent TEVAR. The baseline characteristics of all patients are outlined in This study involved 74 patients with emergent TEVAR including 48 (64.8%) patients with complicated spontaneous AAS (csAAS), including complicated typical TBD (cTBD) . The mean prostheses diameter was 33.1 ± 5.2 mm, whereas the length was 161.7 ± 27.1 mm, respectively. Single stent grafts were used in 70 cases; two stents in 4 patients. In 38 cases, the proximal graft landing zone was below the orifice of the left subclavian artery (LZ 3 and 4), in the other 34 (45.9%) the prosthesis covered the orifice of the left subclavian artery (LZ 2). In 2 subjects operated on in the hybrid room, the landing zone was more proximal (LZ 1) and it was forced to cover the left common carotid artery. Intraprocedural data with a 95.9% primary technical success rate are presented in detail in In the majority of patients, Zenith stent grafts  were implanted. In one case Jotec  and in the other four GORE  stent grafts were used . During in-hospital stays in 3 cases, endoleak type I was reported and the primary clinical success occurred in a final rate of 94.5% [One of the patients died during an in-hospital stay as a consequence of multiorgan failure  on the 19th postoperative day. The patient was operated on due to aorto-oesophageal fistulas but developed sepsis that eventually led to fatal MOF and death. The second patient after primary TEAR presented a severe stroke, never left the hospital, and died 46 days after the procedure. The TEVAR related deaths in total was 2.6% .n = 1, 1.3%), 2 individuals strokes (2.6%) and in 3 cases type I endoleaks were noted. The total prevalence of early neurological adverse events was 4.0%. In one case we observed hemodynamically nonsignificant endoleak and he was discharged in good clinical status and remained asymptomatic, and with a stable view in the follow-up CTA examinations, two next patients were qualified for reintervention. In 18 cases there was a necessity to put the drain to the left pleural cavity, but a few days after TEVAR. Notably, preprocedural bleedings were the predominant reasons. Fifty percent of patients experienced Velazquez postimplantation syndrome (PIS) in the early postprocedural period. No other complications associated with the implantation of the stent graft during the perioperative period were noticed. In our group, the median length of stay in the intensive care unit (ICU) was 1 day, whereas the total hospitalization time was 6 days.Early after procedures, one patient developed transient paraparesis (p = 0.001) , 11 patients died. Annual, five- and ten-year probability of overall survival was 86.4 ± 0.04%, 80.0 ± 0.05%, and 76.6 ± 0.06%, respectively. However, the rate of 5-year survivors was significantly higher after TAI (95.2 ± 3.2%) than csAAS (63.4 ± 5.9%)  [p=0.018), even despite the higher risk profile of the TEVAR group [Patients with complicated TBD require immediate invasive management to prevent death or injury from rupture or organ malperfusion. The IRAD  register confirmed that in 25% of acute B dissection the course is complicated . Trimarc=0.0003) . An alte=0.0003) , 17. The=0.0003) , 19. Adv=0.0003) . Our resAR group . Data frAR group . Patel eAR group . In addiAR group .Endovascular interventions are associated with a risk of serious complications. The most important are endoleaks, stent graft displacement, neurological complications, and retrograde aortic dissection –27. In oOne of the most serious complications following TEVAR procedures are endoleaks, considered by some authors as a failure of the implantation procedure resulting from incorrect planning. Tokyo Consensus indications emphasizes attention on the length of the LZ, aortic angulation, and calcification , 33, 34.The challenging and difficult-to-treat complications are rare erosion of the oesophagus  or the left main bronchus . Takeno In 2002, Fillinger et al. proposed the division of the aorta into five landing zones (LZ) to enable proper planning of the TEVAR strategy, including also hybrid treatment , 33, 34.We are aware that the current study has several limitations. First, the retrospective nonrandomized design and analysis of a limited number of patients from a single center reduces the statistical power of the study. The mortality and morbidity rate are very low as well as general outcomes favorable. Therefore, they may not reflect the results of the other centers. Moreover, we must stress that this report was focused predominantly on mortality and morbidity but the quality of life of survivors was not estimated. The latter aspect would increase markedly the significance of this study; therefore, we plan to conduct such a study in the very close future.Descending aortic pathologies requiring emergent interventions can be treated by endovascular techniques with optimal results and low morbidity and mortality in an experienced and dedicated team. They are characterized by substantially declined mortality and morbidity, notable including also patients with severe clinical status. Key for primary and secondary clinical success is proper technique planning including minimal distal and proximal Landing Zone strategy.


Cyclic filtration tests using bacteria highlighted the competitive antifouling properties of the membranes with a flux recovery ratio after two water/bacterial solution cycles higher than 70%, a reversible flux decline ratio of about 62% and an irreversible flux decline ratio of 28%. Finally, these green antifouling membranes were shown to be stable despite several weeks of immersion in water.While green solvents are being implemented in the fabrication of polyvinylidene fluoride (PVDF) membranes, most are not compatible with the vapor-induced phase separation (VIPS) process for which relatively low dissolution temperatures are required. Additionally, preparing antifouling green membranes in one step by blending the polymer with an antifouling material before inducing phase separation remains extremely challenging due to the solubility issues. Here, the green solvent triethyl phosphate (TEP) was used to solubilize both PVDF and a copolymer (synthesized from styrene monomer and poly(ethylene glycol) methyl ether methacrylate). VIPS was then used, yielding symmetric bi-continuous microfiltration membranes. For a 2 wt% copolymer content in the casting solution, the corresponding membrane P2 showed a homogeneous and dense surface distribution of the copolymer, resulting in a high hydration capacity (>900 mg/cm The extensive use of toxic solvents during membrane fabrication challenges the claim that membrane filtration is a green separation technology. Additionally, as more stringent regulations are being implemented in the utilization of solvents, membrane scientists have started to search for more suitable alternatives and shifted their focus towards membrane fabrication using green or greener solvents ,2,3,4,5.The vapor-induced phase separation (VIPS) process is an adequate process to prepare polyvinylidene fluoride (PVDF) membranes in the microfiltration range , with eir-EGMA). This copolymer was reported earlier as an effective material for the formation of antifouling membranes using traditional (yet toxic) solvents [To answer this need, PVDF was blended in TEP together with a random copolymer synthesized from styrene monomer and poly(ethylene glycol) methyl ether methacrylate (PEGMA), referred to as P with an average molecular weight of 1,500,000 g/mol was obtained from Arkema . It was rinsed with successive baths of deionized (DI) water and methanol before use. Triethyl phosphate (TEP) was purchased from Tokyo Chemical Industry and used without any further purification. DI water was produced in our laboratory with a Purelab® water purification system obtained from Elga-Veolia. Escherichia coli was purchased from the Hsinchu Bioresource Collection and Research Center , while whole blood was obtained from a pool of healthy volunteers at the Taipei MacKay Memorial Hospital . Bovine serum albumin (BSA) was purchased from Sigma-Aldrich .Styrene was bought from Showa Co. PEGMA (Mn 500), and toluene and hexane were purchased from Sigma-Aldrich. Azobisisobutyronitrile (AIBN) was obtained from Alfa Aesar Co.  Poly(vinylidene difluoride)  of the copolymer from the peaks centered at δ = 7.1 ppm (5 protons labeled Hc of the aromatic group of styrene units) and at δ = 3.3 ppm (3 protons labeled Ha of the methoxy group of EGMA units). The FTIR analysis performed with a Jasco FT/IR-6700 spectrometer confirmed the presence of C–C=C  brought stretch ) and C–O stretch ) of PEGMr-EGMA) copolymer and PVDF in TEP under constant stirring. The copolymer content varied between 0 and 2 wt.%, while the PVDF content was fixed at 12 wt.%. The dissolution temperature was fixed at 60 °C. Once the polymers dissolved and the solution was homogeneous, stirring was stopped to allow degassing.Casting solutions were prepared by dissolving P using parallel plate geometry with a diameter of 20 mm (Peltier plate steel). The polymer solution was placed in between the two plates and the gap size set to 100 µm. A 10 s pre-shear at a maximum shear rate of 200 sTM Nano S particle analyzer  after diluting the polymer/solvent or polymer/copolymer/solvent solutions 10 times using TEP.Dynamic light scattering tests were conducted at 25 °C with a DelsaThe membranes were prepared by the VIPS process in a chamber wherein the relative humidity and the temperature were fixed to 70% and 30 °C, respectively. At this temperature, the systems remained homogeneous (no visible phase separation nor gelation occurred). The solutions were hand-cast on glass substrates using a metallic casting knife with a clearance of 300 µm. After 20 min of exposure time to water vapors, the films were immersed in a bath of DI water, dried under atmospheric conditions and then stored in a refrigerator until use.r-EGMA) copolymer and the following number its content in the solution from which the membranes were prepared. Therefore, P0 represents the control virgin membrane (no copolymer).In this work, the membranes were labeled P0, P0.5, P1, P1.5 and P2, where P represents the P(S-r-EGMA)/TEP/water pseudo-ternary phase diagrams were determined by the cloud point method. Briefly, this method consists of preparing a set of polymeric solutions with varying PVDF contents and then adding water dropwise under constant stirring. Once the system becomes permanently cloudy, the cloud point has been reached, and the corresponding composition can be reported in the diagram. The set of obtained compositions at the cloud point is an experimental evaluation of the binodal line. In the case of the pseudo-ternary phase diagram, the PVDF to P(S-r-EGMA) weight ratio was fixed at 6:1 (corresponding to the composition of the casting solution used to prepare the P2 membrane). All phase diagrams were determined at 60 °C, similar to the temperature at which the casting solutions were prepared.PVDF/TEP/water ternary and PVDF/P. Samples were sputter-coated with gold before observation. The membrane pore size was assessed with a capillary flow porometer (PMI). The membrane porosity was determined by gravimetric measurements using butanol. The formula used for the porosity has been reported elsewhere .−1, and 32 scans/spectrum were acquired. For mapping tests, the aperture was 30 µm.The surface chemistry of membranes was analyzed by FTIR and by mapping FTIR, using Jasco equipment . The resolution of each analysis was 4 cmThe water contact angle (WCA) in the air was determined with an optical contact angle system . The hydration capacity was measured on 1.3 cm diameter disk membrane samples. For this, the membranes were dried, weighed and then immersed in DI water for 24 h. Afterwards, they were weighed again and the difference between the wet and dry weights per unit surface area taken as the hydration capacity of the sample. For the WCA and hydration capacity measurements, 5 independent tests were carried out.Escherichia coli bacteria, whole blood and BSA. In all cases, 1.3 cm diameter membrane samples were incubated with the biofouling solution at 37 °C after washing with a PBS solution, and 5 independent tests were performed.The antifouling properties of the membranes in the static condition were tested using Escherichia coli solution prepared according to a procedure earlier reported [® software . In the case of the BSA adsorption tests, the amount of adsorbed protein was evaluated by running a UV–Vis spectrophotometry analysis at 280 nm on the incubation solutions.Regarding the bacterial attachment tests, the samples were incubated for 24 h with 1 mL of reported , and theEscherichia coli solution cycles were performed to evaluate the antifouling properties of the MF membranes during filtration. Firstly, 5 cm disk membranes were immersed in ethanol and placed in a dead-end filtration cell. Then, DI water was filtrated at a pressure of 1.5 bar. After 30 min, the pressure was decreased to 1 bar, the operating pressure used for the test. Once a steady state was reached, the permeate weight was monitored for 30 min. Then, the DI water tank was disconnected from the module and replaced by a tank containing the bacterial solution  and permeate weight monitored for another 30 min. Afterwards, the membranes were cleaned by a backflush procedure at 1 bar for 30 min to remove the reversible fouling. Then, a second DI water/Escherichia coli solution cycle was conducted, followed by a second membrane cleansing and the final DI water permeability measured for the last 30 min of the test. In this study, the flux recovery ratio  to the peak at 1720–1740 cm−1 (P(S-r-EGMA) copolymer).The stability of the membranes was evaluated by immersing the membranes for several weeks in DI water. Then, mapping FTIR (same instrument as mentioned in r-EGMA) destabilizes the system—that is, switches the cloud point curve towards the polymer–solvent axis (r-PEGMA), was added to the systems. Finally, the retainment of the bi-continuous structure after phase inversion by SD is facilitated, because all solutions can readily gel . It means that the structure cannot evolve (or hardly) after phase inversion, which would have led otherwise to the formation of cellular domains after coalescence of the polymer chains.In this study, large membranes could be hand-cast. The resulting films visually seemed homogeneous, whitish and matte . These vent axis b. This aent axis c becauser-PEGMA (2 wt%). As seen earlier from the SEM images of Finally, the pore size and porosity were also determined and the results shown in −1 corresponding to the C=O stretch of the PEGMA moieties. It could also be identified on the ATR-FTIR spectra of the membranes shown in −1  copolymer are also effective in reducing biofouling in porous membranes despite their propensity to promote the physical attachment of bacteria on surface pores, providing a homogeneous and dense distribution of the antifouling units on the surface  copolymer plays a similar role and significantly contributes to mitigating blood cell attachment on the membrane, and a 90% reduction in cell adhesion was measured with the P2 sample. It implies that the as-prepared green membranes improved the hemocompatibility compared to the unmodified membrane, which would be essential for application in biomedical devices.As mentioned earlier, the structures of the prepared membranes make them suitable for rejecting large biofoulants. Blood cells also fall into this category, and MF membranes can be used in biomedical applications in contact with blood, such as, for example, in leukodepletion . Interac−1 (amide group of BSA [2 vs. 82 µg/cm2 for the virgin membrane). Compared to the results of previous sections in which larger biofoulants were used, it is undeniable that more fouling occurred. Nevertheless, it remained low and comparable with the adsorption level measured by Chiag et al. [Proteins foul membranes at a lower scale than cells, as their small size facilitates their entrapment in the porous structure. BSA is commonly employed as a model protein to study the fouling of porous membranes in adsorption tests ,46,47 orp of BSA ) to asseg et al. . InteresEscherichia. coli was used as the model, as it is relatively safe to handle.In order to demonstrate the reduced fouling propensity on the green membranes during operation, cyclic water/bacterial solution filtration tests were conducted. Bacteria were chosen, as these are large biofoulants, and VIPS membranes would normally be used to reject particles or microorganisms of similar size. Additionally, Although the prime purpose of the test was to study biofouling during operation and not to develop membranes for the removal of bacteria from wastewater, we still determined the rejection. It was 86 ± 3% for the virgin membrane and 97 ± 2% for P2. These rejections may seem low considering the membrane pore size and that of the bacteria (rod-shaped bacteria of a length of about 2 µm and diameter 0.5 µm). However, it is known that the cell wall of Gram-negative bacteria gives them flexibility. As such, they can deform and squeeze through smaller pores than their size under a pressure gradient , preventThe permeate flux was recorded and is shown in One major and commonly reported drawback of hydrophilic modifiers for membranes is that they can leach out of membranes, due to poor stabilizing interactions with the hydrophobic membrane material ,56,57,58r-EGMA) copolymer, an antifouling copolymer. Relatively low temperatures (60 °C) could be used to prepare the casting solutions, hence demonstrating that this green solvent is suitable for the formation of antifouling membranes in one step by VIPS or by other phase inversion processes that require low dissolution temperatures. The membranes obtained were bi-continuous, highly porous and with a homogeneous surface distribution of the copolymer. Their ability to trap a large amount of water resulted in the efficient mitigation of biofouling tested with various biofoulants  in static and dynamic conditions. In addition, the membranes remained stable even after a long immersion in DI water (>98% copolymer remaining after 4 weeks). All in all, this work proves that green antifouling membranes can be prepared by VIPS, despite the scarcity of green solvents suitable for nonsolvent-induced phase inversion processes. The formation of green PVDF membranes by the VIPS process using zwitterionic copolymers is also under investigation by our team, considering the outstanding antifouling performances of these materials.This work presented the formation of antifouling and green microfiltration membranes by the VIPS process. TEP was employed as a green solvent to form homogeneous polymeric systems containing PVDF, the main membrane polymer, and P(S-


Phakopsora pachyrhizi) in the southeastern USA.Eight soybean genomic regions, including six never before reported, were found to be associated with resistance to soybean rust (Phakopsora pachyrhizi is one of the most important foliar diseases of soybean [Glycine max (L.) Merr.]. Although seven Rpp resistance gene loci have been reported, extensive pathotype variation in and among fungal populations increases the importance of identifying additional genes and loci associated with rust resistance. One hundred and ninety-one soybean plant introductions from Japan, Indonesia and Vietnam, and 65 plant introductions from other countries were screened for resistance to P. pachyrhizi under field conditions in the southeastern USA between 2008 and 2015. The results indicated that 84, 69, and 49% of the accessions from southern Japan, Vietnam or central Indonesia, respectively, had negative BLUP values, indicating less disease than the panel mean. A genome-wide association analysis using SoySNP50K Infinium BeadChip data identified eight genomic regions on seven chromosomes associated with SBR resistance, including previously unreported regions of Chromosomes 1, 4, 6, 9, 13, and 15, in addition to the locations of the Rpp3 and Rpp6 loci. The six unreported genomic regions might contain novel Rpp loci. The identification of additional sources of rust resistance and associated genomic regions will further efforts to develop soybean cultivars with broad and durable resistance to soybean rust in the southern USA.Soybean rust caused by The online version contains supplementary material available at 10.1007/s00122-022-04168-y. Phakopsora pachyrhizi Syd. is one of the most important foliar diseases of soybean [Glycine max (L.) Merr.]. This obligate biotrophic fungus has spread from East Asia to every continent where soybean is grown. After becoming established in South America, it was first observed in the continental USA in November 2004  caused by G. max and other Glycine species, including the perennial species G. tomentella  6 and that 37 of the 52 PIs had a SNP haplotype identical to that of PI 462312, the accession in which the Rpp3 locus was first discovered. The high percentage of PIs with a resistance allele at one of these loci limits the number of potential resistance gene pyramids, or combinations of Rpp genes that could provide broader and more durable resistance  have been reported to be resistant to P. pachyrhizi isolates from Thailand, Brazil, Paraguay, and Zimbabwe. Seedling assays were conducted in a biosafety level 3 greenhouse, and based on the criteria used to define resistance, the authors concluded that 805 accessions (< 4.9%) had resistance to the four isolates. When the 805 PIs were tested in the field between 2006 and 2008, however, only 64 (8%) were confirmed to be resistant to seven southern US populations of the rust fungus , Phytophthora sojae, and other soybean pathogens  to search for significant genotype–phenotype associations using single nucleotide polymorphism (SNP) markers have been successfully used to understand genetic architecture in panels of germplasm lines and to identify regions of the soybean genome associated with phenotypic variation in traits like resistance to soybean cyst nematode (P. pachyrhizi under in the southeastern USA and (2) identify genomic regions associated with soybean rust resistance in those accessions, many of which had been reported to be susceptible to four foreign isolates. Germplasm accessions from a variety of countries and regions were screened initially, but screening was later focused primarily on PIs from Vietnam, southern Japan, or central Indonesia after preliminary evaluations showed that most of the accessions with resistance originated from those three geographic regions.Objectives of this study were to (1) evaluate soybean germplasm accessions for resistance to field populations of Rpp genes at known loci  than the tests planted in subsequent years. After 2008, most of the accessions screened originated from southern Japan, Vietnam, or central Indonesia, and were in MGs IV through IX. This was because data from the earlier screenings had shown that few PIs from any other regions were resistant in the southern USA. Seeds were obtained from the USDA Soybean Germplasm Collection in Urbana, IL, USA.Two hundred and fifty-six germplasm accessions from Japan, Indonesia, Vietnam, China and several other countries were screened for their reactions to SBR in multiple year-location field tests, along with susceptible controls and PIs with i Tables  and S1. i Tables . After tRpp genes were generally planted in each test also to permit detection of possible pathotype differences among P. pachyrhizi populations in different years and locations. PIs carrying the Rpp1 through Rpp6 genes were planted, including Rpp1, Rpp2 and Rpp5 accessions with different resistance alleles. The Japanese cultivar Hyuuga (PI 506764), which has resistance alleles at the Rpp3 and Rpp5 loci , as described in Walker et al. . SBR disease severity was rated either on plants in the field, or on leaflets collected from test plots between mid-October and early December, when SBR symptoms and signs  were visible on susceptible checks. At least 10 leaflets were collected randomly from the mid-canopy, placed in labeled bags, and kept in a refrigerator until they could be examined with a compound microscope. Reactions to SBR in some test locations were evaluated using different rating methods and scales in 2008, when tests were grown in four different states. The scales and methods used at each location between 2008 and 2011 were described in Walker et al. . The 20 accessions with the lowest disease ratings  were labeled in the PC plot to facilitate the assessment of the importance of geographic sources of resistance in the panel. Accession names in the PCoA plot and neighbor joining tree were colored based on the country of origin of each PI to make visual assessment more informative.To assess the degree of population structure, principal coordinate analysis (PCoA) was conducted in the GAPIT Version 3 R package , which permitted us to overcome the limitations of an incomplete dataset and the differences among test environments. BLUP values were calculated by treating genotype, environment, and genotype × environment interactions as random variables using the Standard Least Squares and restricted maximum-likelihood (REML) methods Shaw . The BLUp < 1.445 × 10–5) were considered significant.A GWAS analysis was conducted using the Fixed and random model Circulating Probability Unification (FarmCPU) method  and PI 567102B (Rpp6) were typically between 1.0 and 2.0, and BLUP values less than − 1.00. Differences in the reactions of some accessions with known Rpp genes between test environments indicated pathotype variation among some of the P. pachyrhizi populations.SBR severity and urediniospore production on susceptible checks demonstrated that disease pressure was generally high by the time the PI reactions were rated in each test. Rust index (RI) ratings for susceptible checks were between 3.5 and 5.0 in most tests, resulting in positive BLUP values that were often higher than 1.00 Table . In contRpp6), PI 200492 (Rpp1), PI 567090, PI 567102B (Rpp6), PI 200532, PI 567061, PI 635999, PI 605823 (Rpp7) and PI 200547 , and 24 were from northern Vietnam. Among the few resistant accessions not from those areas were PI 518295 from Taiwan and PI 476905A from an unknown location in China. Of the other 23 accessions from China, only PI 594796 and PI 594742 had negative BLUP values, and those were only − 0.07 and − 0.01, respectively (Table S1).BLUP values calculated from the disease ratings ranged from − 1.46 (least disease) to 1.60 (most disease). One hundred and thirty-eight accessions had BLUP values of − 0.10 or lower, and the seven most resistant accessions had BLUPs less than − 1.00 Tables  and S1. Rpp3 gene in PI 462312, the allele(s) of Rpp2 in PI 230,970 and PI 417125, the Rpp5 alleles of PI 200487 and PI 471904, and at least one of the two Rpp genes in ‘Hyuuga’ (PI 506764)  of PI 594760B and PI 594767A did not protect plants against the pathotypes encountered. The Rpp3 allele in PI 567099A was also ineffective compared to the Rpp3 gene in PI 462312. The Rpp5 allele(s) in PI 200487 from southern Japan and in PI 471904 from East Java provided resistance, whereas the Rpp5 allele from PI 200526 and the recessive rpp5 allele from PI 200456 did not suppress disease in those two Japanese accessions (Table S1). The Rpp4 allele that Harris et al.  in PI 567102B and PI 567104B. At least five PIs with single Rpp genes had less disease than the cultivar Hyuuga (PI 506764), which has resistance alleles at the Rpp5 and Rpp3 loci  Fig.  and the P < 1.445 × 10–5; − log10(P) = 4.8)  values, in descending order of probability, were on Chrs 4, 9, 15, 13, 1, and 18  the majority of the resistant accessions originated from either Japan, Vietnam or Indonesia; and (3) GWAS analysis of BLUP values calculated from disease reactions in multiple years and locations could be used to identify eight genomic regions putatively associated with resistance to SBR. Six of those regions have not previously been reported as far as we know.This study demonstrated that (1) numerous soybean PIs previously reported to be susceptible to Rpp genes and genomic regions of the soybean genome associated with resistance to SBR have been identified in previous studies using biparental molecular mapping, bulked segregant analysis or GWAS with seedling reaction data from greenhouse assays , central Indonesia (Rpp6), or Vietnam (Rpp7). Although PI 462312 (‘Ankur’), the source of the Rpp3 gene, was selected in Uttar Pradesh in north-central India, it was selected from a cross between unknown parents made in the USA, so the original source of the resistance gene is not known (Germplasm Resources Information Network). In this study, 84% of the accessions from Japan, 69% of those from Vietnam, and 49% of those from Indonesia had negative BLUP values, compared to only 11% of the 27 Chinese PIs tested. Nevertheless, some PIs from Japan, Vietnam or central Indonesia were susceptible in our tests; the Japanese accessions PI 200456, PI 200526 (Shiranui) and PI 224270 (Hougyoku) had positive BLUP values, even though Yamanaka et al. , which had an RB reaction in Hanoi, and PI 459025B (Rpp4), which had an intermediate reaction there. In contrast, PI 200492 (Rpp1), PI 462312 (Rpp3) and PI 417089A  were resistant in our tests but developed TAN infection types in Hanoi  were resistant there. These results attest to the pathogenic diversity among P. pachyrhizi populations.PI 635999 (‘DT 2000’) and PI 423972 were also resistant in field studies conducted in Hanoi, Vietnam, in 2006 and 2007 . Six of 24 Chinese PIs screened are known or thought to have a resistance gene at the Rpp1 locus, but like the Rpp1-b gene of PI 594538A and the Rpp1 allele from PI 561356, none of their alleles provided resistance in the southern USA , Vietnam (PI 635999) and several from Japan, such as PI 416826A and PI 200488. Hyuuga and PI 462312 had similar BLUP means (− 0.64 and − 0.66), suggesting that the Rpp5 allele in Hyuuga may not have enhanced resistance against US rust populations. Resistance genes at the Rpp6 locus on Chr 18 appear to be less common, but the allele from PI 567102B and PI 567104B conditioned very high levels of resistance, resulting in the lowest and fourth lowest BLUP means. PI 567090, which had the third lowest BLUP value, has a resistance allele at the Rpp3 locus and another on Chr 18, likely at the Rpp6 locus  was the only highly resistant accession from the present study that also had resistance to most of the 16 isolates. These results and those of Pham et al.  were effective. Of the eight genomic regions that were significant at P < 1.445 × 10–5; − log10(P) = 4.8, only ss715594707 on Chr 6 and ss715632525 on Chr 18 were locations with known Rpp loci. The former is near the Rpp3 locus, a common location for genes that condition resistance to SBR in the USA  are known or thought to have an SBR resistance gene at the Rpp3 locus  in seven Chinese accessions were ineffective against the US field pathotypes. As a result, the frequency of Rpp1 alleles with significant phenotypic effects in this study was low, which probably explains the reason that the locus was not detected using GWAS. Other than PI 200492, PIs 417120 and 423958 from Japan, and PI 518295 from Taiwan were among the few accessions screened that had an effective resistance allele at the Rpp1 locus .When Chang et al.  performeRpp2 on Chr 16;  and PI 200487  were among the 20 accessions with the lowest BLUP means for disease in this study, Rpp7 has not been reported in any other accessions, and PI 471904 appears to be the only other PI with an effective allele at the Rpp5 locus. A failure to detect genes that are present at a low frequency in a population is one limitation of GWAS analyses Below is the link to the electronic supplementary material.


Reverse periodization is commonly touted as a salient planning strategy to improve sport performance in athletes, but benefits have not been clearly described.We sought to identify the main characteristics of reverse periodization, and the influence of training volume and periodization models on enhancing physiological measures and sports performance.Systematic review.The electronic databases Scopus, PubMed and Web of Science were searched using a comprehensive list of relevant terms.A total of 925 studies were identified, and after removal of duplicates and studies based on title and abstract screening, 17 studies remained, and 11 finally included in the systematic review. There was a total of 200 athletes in the included studies. Reverse periodization does not provide superior performance improvements in swimming, running, muscular endurance, maximum strength, or maximal oxygen uptake, compared to traditional or block periodization. The quality of evidence levels for the reverse periodization studies was 1b  for two investigations, 2b  for the remaining studies and a mean of 4.9 points in the PEDro scale (range 0–7).It appears that reverse periodization is no more effective than other forms of periodization in improving sports performance. More comparative studies on this alternative version of periodization are required to verify its effectiveness and utility across a range of endurance sports. Reverse periodization is no more effective than other forms of periodization in improving sports performance, muscular endurance, maximum strength, or maximal oxygen uptake.The use of reverse periodization likely induces similar improvements to a traditional model in shorter events such as the 100-m swimming event.More comparative studies of periodization models in endurance sports require careful planning of experimental design, longer study periods, and where appropriate matching of training volumes and intensities.Periodization is a process that serves as the macromanagement of an athlete’s training program in the context of the annual plan , 2. MatvThe rationale of periodized models of strength and power training in athletes originated in western countries centering on the work of Stone and O´Bryant , Stone apolarized three zone model of training is another approach characterized by covering ~ 80% of the volume in zone 1 (blood lactate [La−]b ≤ 2 mmol L−1) with most of the remaining 20% conducted in zone 3 (above velocity of 4 mmol L−1) [Over recent decades, many approaches have evolved that can be broadly categorized as traditional, block, or reverse periodization, each offering a differing rationale and template for subdivision of the training program into sequential elements. Bompa  classifimol L−1) , 33. Revmol L−1) , 34, modmol L−1) , 35, recmol L−1)  and femamol L−1) .All periodized models  can be considered a useful means of coordinating training to improve human sporting performance. However, more research is needed to provide a better understanding of the benefits of reverse training periodization in comparison with other models. The aim of this study was to conduct a systematic review of periodization studies to evaluate the effectiveness and utility of reverse periodization, and the influence of training volume/intensity in enhancing sports performance.A literature search was completed in December 2021 by two independent researchers (VR-C and JM-G) using the three industry-standard databases with no date restrictions: PubMed, Web of Science and Scopus. The search strategy consisted of identifying the relevant studies, with all terms searched in the title, abstract and keywords (where applicable).This systematic review was conducted following the guidelines of the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement .The keywords used in the searches were: periodization, training, reverse, linear traditional and block. Title, abstract and keyword fields were searched using the following search strategy: ((("periodization" OR "Training") AND "reverse") AND ).Following the literature search, the identification, screening, eligibility assessments and inclusion of studies were performed by the same researchers with disagreement settled by consensus. All duplicate references were removed, and remaining records examined by title and abstract to exclude irrelevant records. Studies were then selected following the eligibility criteria Table . DescripThe summary of eligibility criteria is shown in Table The level of the sample was classified as recreational and trained athletes using the criteria of each study included in the systematic review.Two of the authors (VR-C and JG-R) independently extracted characteristics of training protocols and results using a standardized form. A total of 11 studies were identified  analyzed the quality of included studies using the modified PEDro scale  and OxfoA total of 925 potential manuscripts were identified following database examination Fig. . ReferenIn terms of the quality of the studies selected, all studies were evaluated with the PEDro scale, with a mean score of 4.91 Table . Using tSix studies were conducted in mostly recreational athletes and five in trained athletes. There were a total of 230 athletes in the included studies, involving a total of 134 females (58%). The mean age of the athletes was 23 y (standard deviation of 6 y), with a range of 16–37 y. Two of the studies assessed females only, nine studies involved both males and females, and none of the studies assessed males only. In addition, only two studies used a control group to evaluate periodization models during the experimental intervention. The training programs evaluated in this review were predominantly short-term interventions , 41, 43 Three typical patterns detailing the distribution of training intensity in a macrocycle - traditional periodization, block periodization and reverse periodization - are illustrated in Fig. vs. 160 km). In addition, the traditional periodization group performed 324 km compared to 212 km for the reverse periodization group in the study of Arroyo-Toledo et al. [Regarding training volume, the running studies reported the volume using different metrics of either time or distance. The athletes in the study of Gómez Martin et al.  performeo et al. . Howevero et al. . Finallyo et al.  performeo et al.  lifted b2max), (ii) lactate threshold and (iii) movement economy. Both reverse periodization and block periodization training have yielded similar improvements in V̇O2max and the velocity corresponding to V̇O2max (vV̇O2max) [2max for reverse periodization and reductions for the traditional periodization model were reported in the study of Clemente-Suárez et al. [2peak) were reported for traditional and reverse periodization [There are three main physiological parameters  affectinV̇O2max) . Greaterz et al. . Similardization . Energy dization . Finallydization .Two studies , 31 repon = 9) and block periodization (n = 2) training with reverse periodization. Studies were conducted in both recreational [This systematic review identified 11 studies that directly compared traditional periodization  performance, 5 of the 11 studies included in this review were in swimming. A systematic review on swimming periodization identified that the traditional periodization was the most common form used in well-trained swimmers, but only four studies compared traditional versus reverse periodization  more thaswimmers . Reverseswimmers , 43, witswimmers . Howevervs. reverse periodization) in bench press (17% and 16%), lat pull-down (30% and 22%), arm curl (20% and 16%) and leg extension (37% and 32%). However, Prestes et al. [To our knowledge, only two studies have reported greater gains in 1RM strength in traditional periodization/programming as opposed to reverse periodization/programming , 42. Regs et al. , assertes et al. . A simils et al. . Althougs et al.  formallyIn addition to effects on performance and physiological parameters, different types of periodization may have variable effects on body composition. Arroyo Toledo et al.  reportedThere were some limitations to this review given the heterogeneity of sports, training and methodological approaches of the underlying studies. There was substantial inter-individual variability regarding the participants in the different studies  across all performance variables that may have impaired the ability to establish conclusive outcomes in this systematic review. In addition, as periodization generally refers to periods of a season or more, it may be logical for future research to evaluate longer periods, so that differences after each periodization model can become more pronounced. A critical drawback in some of these studies is the lack of a randomized controlled design  as shown in Table It is not clear if reverse periodization is more effective in improving sports performance than other periodized models. Use of reverse periodization likely induces similar improvements to a traditional model in shorter events such as the 100-m swimming event. Comparative studies of periodization models in endurance sports require careful planning of experimental design, longer study periods, and where appropriate careful matching of training volumes and intensities.


Prevotella, Cetobacterium, and Megamonas was observed in patients with HFMD, whereas a large increase in genus Bacteroides, Ruminococcus, and Faecalibacterium were seen in the control group. We also share the bioinformatic analytical pipeline for this analysis, from data preprocessing to data filtering and amplicon sequence variant (ASV) table generation. We expect that the dataset will be reprocessed, evaluated and fully analysed with various analysis methods to further elucidate the role of the gut microbiota in HFMD development.There is evidence linking hand, foot and mouth disease (HFMD) to gut microbiota dysbiosis, and this relationship was corroborated in a large HFMD patient population in our previous study. Here, we present a bacterial 16S rRNA gene dataset from faecal samples of 713 individuals  aged 2 to 7 years residing in Heyuan and Jiangmen counties, Guangdong Province, southern China. Microbiome analysis indicated a significant increase in genus  Microorganisms are highly dynamic and variable among individuals and throughout development due to environmental and individual factors, such as host genotype, diet and lifestyle5. With advances in high-throughput sequencing technologies and related bioinformatics methods, especially 16S ribosomal RNA (rRNA) gene analysis, we have been able to obtain deeper insight into the effects of intestinal microorganisms. 16S rRNA gene analysis has become the main focus of meta-taxonomic analysis and is widely used in microbiome research7.Thousands of microorganisms colonize the gastrointestinal (GI) tract and work in concert with the intestinal epithelial barrier to defend against pathogen invasion8. The predominant clinical manifestations of HFMD are a transient maculopapular rash and blisters or ulcers involving the hands, soles of the feet and mouth that last approximately half a week with no observable sequelae in most cases9. The specific pathogenesis of HFMD is not completely understood.Hand, foot, and mouth disease (HFMD) is a self-limiting disease resulting from infection with some human enteroviruses, including enterovirus A71 (EV71), coxsackievirus A16 (CVA16) and coxsackievirus A10 (CV-A10), that is prevalent in the neonatal and paediatric populations11. In addition to the beneficial effects of bacteria in the gut, the significance of the gut bacterial composition in the emergence and development of certain diseases has been emphasized in some studies. The presence of harmful intestinal bacteria may induce the development of inflammatory bowel disease12. The intestinal microbiota of individuals with HFMD has been demonstrated to be altered in type and relative abundance of bacteria, and these alterations correlate with the severity of HFMD13. Whether a similar relationship exists between gut flora dysbiosis and the occurrence of HFMD is unknown. Only limited studies with very small sample sizes have focused on characteristic changes in the gut microbiota profile of children with HFMD. To further reveal the role and underlying mechanisms of intestinal flora dysbiosis in the pathogenesis of HFMD, larger population studies are needed.Although enteroviruses are the causative agents of HFMD in children, the role of the gut microbiota, constituting the intestinal microorganisms, in enterovirus pathogenesis and disease progression needs to be considered. The gut microbiota is a key component in intestinal immune responses and repairing the injured intestinal mucosa barrier to protect against pathogen invasion and maintain homeostasisPrevotella and Streptococcus were increased in the case group, while the relative abundances of the beneficial bacteria Bifidobacteria and Faecalibacterium were decreased. In addition, we concluded that administered synbiotics supplements potentially provide better resistance to gut dysbiosis in patients with HFMD than probiotics or prebiotics alone. Greater gut flora disturbance was observed in patients who were enterovirus A71 (EV71)-positive than in those who were coxsackievirus A16 (CAV16)-positive14.Guangdong Province, an area with a high prevalence of HFMD in the Asia-Pacific region, has several designated HFMD surveillance sites, including Heyuan City and Jiangmen City. In a previous case‒control study, we enrolled 749 participants  from the Heyuan and Jiangmen regions and obtained faecal samples for 16S rRNA gene sequencing, followed by bioinformatics analysis for profiling. We compared the composition and variability in gut microbe communities between the different groups, among different enterovirus types and according to supplement intake. The results showed that the distribution characteristics of the intestinal flora in individuals with HFMD differed from those in healthy subjects. Compared with those in the healthy group, the relative abundances of We reorganized the database in order to maximize the presentation of our database and to make the dataset of higher quality. The final submitted dataset was from 713 study participants, including 459 healthy children and 254 patients with HFMD. And an analysis result of case-control study in the ratio of 1:1 was also provided. The details pertaining to the study design, sample collection, raw sequence data, bioinformatics analysis pipeline, and results of our bioinformatic analysis are provided in the subsequent sections. We expect the dataset provided here to be fully utilized and comparatively evaluated by the scientific community to provide detailed insight into the potential relationship between the gut microbiota and HFMD; understanding this relationship may contribute to improving intervention and treatment strategies for the disease.All participants were selected from a previous investigation conducted by the Guangdong Institute of Public Health in 2017 that aimed to explore the distribution characteristics of gut microbiota and its related influencing factors in children with HFMD. The previous study was approved by the Ethical Review Board of Guangdong Provincial Center for Disease Control and Prevention (NO: 2016016). The legal guardians/parents of all the children were informed of the study objectives and sampling procedure and provided written informed consent prior to completing the survey. Personal identifiable information pertaining to participants was kept private. The cohort size for the study was statistically derived. Supplementary Fig. Two distinct centres, including Xinhui District of Jiangmen City and Longchuan County of Heyuan City in Guangdong Province were investigation sites and two local hospitals, including a children’s hospital and a county-level hospital, were selected in each investigation site to collected specimens. The case group at each investigation site was randomly selected from the local outpatient departments of hospitals, and the control group was recruited from the same hospitals or the neighbourhood of the patients. The participants were required to complete a disease risk-factor questionnaire that collected information about demographic characteristics, hygienic habits, and dietary habits in the last month. The selection criteria used for study inclusion were as follows: a) age between 2 and 7 years old; b) residency in one county or district for at least 6 months; and c) HFMD diagnosed by medical institutions in the county or district according to the Chinese Guidelines for HFMD Diagnosis and Treatment issued by the Ministry of Health in 2010, and the results were reviewed by the laboratory of Guangdong Provincial Center for Disease Control and Prevention. The exclusion criteria were as follows: a) a history of critical illness, herpetic pharyngitis or severe organic lesions; b) a disease related to the risk factors in the study, including diarrhoea, gastroenteritis, previous diagnosis of HFMD, or herpetic angina, among others; or c) having potential risk of exposure to or taking antibiotics within a month.Participants were required to deliver faecal samples of approximately 5–8 g within 7 days of the onset of illness. At the laboratory, samples were immediately placed into a sterile tube and labelled with the child’s name, identification code and bristol classification number. The collected samples were reserved in a 4 °C refrigerator for temporary preservation within 2 h, stored in a −18 to −20 °C freezer for storage within 12 h and transferred to a −80 °C freezer after 12 h until further processing. All collection staff wore sterilized, clean, new laboratory gloves and facemasks to avoid bacterial contamination in this study.Genomic DNA was extracted with the MagaBio Feces Genomic DNA Purification Kit  following the manufacturer’s instructions. Subsequently, amplicons of the V4-1 hypervariable region of the bacterial 16S rRNA gene were generated using the specific primers 515 F/R806 (GTGCCAGCMGCCGCGGTAA- GGACTACHVGGGTWTCTAAT) with 12-bp barcodes. Primers were synthesized by Invitrogen . A PCR BioRad S1000 instrument  with a PCR mixture containing 25 μl of 2x Premix Taq , and 1 μl of each primer (10 μM) and 3 μl DNA (20 ng/μl) template in a total volume of 50 µl were used for bacterial amplicon amplification. The thermocycling scheme was initialization for 5 min at 94 °C; 30 cycles of denaturation for 30 s at 94 °C, annealing for 30 s at 52 °C, and extension for 30 s at 72 °C; followed by final elongation for 10 min at 72 °C. Only samples with bright primary bands between 290 and 310 bp were used in further experiments. Sequencing libraries were constructed using the NEBNext Ultra II DNA Library Prep Kit for Illumina  according to the manufacturer’s recommendations. Finally, the library was sequenced on the Illumina Nova6000 platform, and 250 bp paired-end reads were produced at Guangdong MagiGene Biotechnology Company .https://github.com/YongxinLiu/EasyAmplicon15. The pipeline allows analysis and visualization of microbiome data, especially 16S rDNA amplicon sequencing. Three main analysis steps of the pipeline were used in this analysis, including sequence data preprocessing, feature table construction and processing, and species diversity and differential analyses. All bioinformatic and statistical analyses were conducted in R software, version 4.1.3  unless otherwise stated. Before raw data processing, the statistics for each sample sequence were computed with the seqkit program16 (v0.15.0), and the results are available in Figshare File 117.A 1:1 matched case-control study among 713 participants was implemented by using SPSS 23.0  to minimize the effect of potentially confounding factors age and sex. 229 patients with HFMD and 229 healthy control subjects reserved after case control matching. Figure 18 (fastq_mergepairs) was used to combine the forward and reverse reads. We identified and removed primer sequences at the beginning and end of the sequence using VSEARCH  to control the maximum error rate within 1% (fastq_maxee_rate 1%). Then, the filtered reads were dereplicated by omitting population-level singletons to identify the set of unique sequences (VSEARCH derep_fulllength: minuniquesize 500). Unique sequences were denoised with the UNOISE algorithm, which controlled the ASV number to reduce the size of the dataset and achieve single-base ASV accuracy (USEARCH v10.0.240 unoise3: minsize 10)19. Finally, the program VSEARCH with reference mode (uchime_ref) mapping against the Ribosomal Database Project database (RDP_16 S_V18)20 was used for chimaera detection and removal.Sequence merging, primer removal, quality filtering, and dereplication and denoising of amplicon sequence variants (ASVs) were performed in the preprocessing step. According to the principle of overlap between paired-end reads, the software VSEARCH version 2.15.217. Eventually, to standardize the sequencing depth among the samples, the analysis with the lowest number of sequences by random resampling  was performed using ‘otutab_rare.Rscripts’ with the ‘vegan’ package (v2.5–7).In the second step of the pipeline, we constructed a feature table of ASVs abundance by mapping all the filtered reads to the list of chimaera-free ASVs, with a similarity threshold of 0.97, using VSEARCH . Additionally, taxonomy was annotated using the ‘VSEARCH–sintax’ command (sintax_cutoff 0.6) in combination with the RDP database . ASV_247 were removed from the dataset using ‘otutab_filter_nonBac.Rscripts’, as those were annotated as unclassified ASVs. The filtered ASVs table are displayed in Figshare File 3et al.21 by using custom code  bacteria in each group was conducted with ‘otu_mean. Rscript’. Linear discriminant analysis effect size (LEfSe) was performed with a Linear Discriminate Analysis (LDA) using ‘format2lefse.Rscript’ to determine the potential taxa that most likely explain the differences between the case and control groups. Finally, we combined the feature tables with annotated species information into tables at the phylum , order , class , family  and genus levels ; such tables allowed further analysis of different levels of gut flora17. In total, raw RDP taxonomic profiles generated a total of 551 ASVs, which were annotated to 8 bacterial phyla, 15 classes, 21 orders, 38 families, and 103 genera.Analyses involving the calculation of alpha diversity indices utilized the Shannon, Chao1, ACE, Richness, Simpson and InvSimpson indices, which are sensitive for high-abundant bacteria and can be computed by using R software with the ‘USEARCH -alpha_div’ command. To capture the contribution of low-abundant bacteria, the Tail statistic, a rank-based diversity measure was calculated as described by Li https://ngdc.cncb.ac.cn/gsa/browse/CRA009110)23. The pipeline used for this analysis can download through Github platform (https://github.com/YongxinLiu/EasyAmplicon). Other data types, such as metadata, some taxonomic abundance datasets, and statistical results are accessible through Figshare platform. Visualizations of the analysis results have been provided as a part of this publication.A 16S rRNA gene raw sequencing dataset entitled 16S rRNA gene sequence data from children with hand, foot and mouth disease and healthy individuals is available for download on the Genome Sequence Archive (GSA) platform, under GSA ID CRA009110 . The purpose of sharing the FASTQ data is to allow other researchers to verify, reprocess and reanalyse the data according to their analytical needs and customized parameters.17 and Figshare File 1017 to the article, respectively.Critical epidemiological information of the study population is included in the metadata to provide a reference for future retrospective analyses by other researchers. The metadata consists of demographic information , type of enterovirus infection, hygienic habits, dietary habits  and information about the inserted primer sequences. Metadata for 713 participants and 458 participants were also provided as Figshare File 9Statistical report of each sample sequence before merging were presented as File 1 in the Figshare. Taxonomy annotation tables for each ASV based on the RDP database can be accessed through File 2. And RDP classified taxonomic profile at five levels of taxonomic lineage is available in the files titled File 4 through 8.The participant inclusion and exclusion criteria were strictly enforced, and the epidemiological information entry and stool collection processes for each subject were checked multiple times to ensure the authenticity and validity of the data.24 software after merging the forward and reverse reads, and the results showed that the quality score of each sample was above the Q20 reported accuracy, allowing further analyses. A total of 34,448,262 reads were identified among the 458 samples, with an average read length of 291.90 bases per sample. Quality control of merged sequences was performed using VSEARCH to control the expected allowed error in the amplicon sequence and obtain paired-end clean tags, resulting in 33,295,761 good quality reads. In addition, 565 ASVs were identified by USEARCH-based de novo classification, and 551 unique ASVs were screened by VSEARCH after removing unclassfied sequence.The quality and concentration of extracted DNA were evaluated using a Thermo NanoDrop One instrument . We verified the amplification length and concentration of the V4-1 region of the 16S rRNA gene by performing 1% agarose gel electrophoresis and visualizing the bands. Library quality was assessed on a Qubit Fluorometer . Raw sequencing data quality was checked using FASTQC v0.11.9Bacteroides, Prevotella, Faecalibacterium, Parabacteroides, and Bifidobacterium were the dominant genera (Fig. Prevotella, Cetobacterium, and Megamonas were enriched in the HFMD group, whereas HFMD-related genera, including Bacteroides, Ruminococcus, and Faecalibacterium were enriched in the control group (Fig. Comparisons of microbiota composition and differences between the case and control groups was conducted based on the feature table. Differences in high-abundant bacteria between the HFMD group and the control group were observed in the alpha diversity analysis based on the Simpson and Shannon indices Fig. . Similarera Fig. . To furtoup Fig. .Fig. 2ChFlow chart of subjects selection along with inclusion and exclusion criteriaThe collection of Supplementary Figure 1 and Supplementary File 1


Leucoraja erinacea) that the pseudobranch does, in fact, derive from the mandibular arch, and that it shares gene expression features and cell types with gills. We also show that the skate mandibular arch pseudobranch is supported by a spiracular cartilage that is patterned by a shh-expressing epithelial signalling centre. This closely parallels the condition seen in the gill arches, where cartilaginous appendages called branchial rays, which support the respiratory lamellae of the gills, are patterned by a shh-expressing gill arch epithelial ridge. Together with similar discoveries in zebrafish, our findings support serial homology of the pseudobranch and gills, and an ancestral origin of gill arch-like anatomical features from the gnathostome mandibular arch.The pseudobranch is a gill-like epithelial elaboration that sits behind the jaw of most fishes. This structure was classically regarded as a vestige of the ancestral gill arch-like condition of the gnathostome jaw. However, more recently, hypotheses of jaw evolution by transformation of a gill arch have been challenged, and the pseudobranch has alternatively been considered a specialised derivative of the second (hyoid) pharyngeal arch. Here, we demonstrate in the skate ( Summary: The skate pseudobranch is a gill serial homologue and reveals the ancestral gill arch-like nature of the jawed vertebrate mandibular arch. Hypotheses of jaw-gill arch serial homology are deeply rooted in the fields of vertebrate evolutionary biology and comparative anatomy, and have profoundly shaped views on the evolutionary origin of the jawed vertebrate body plan. Apparent correspondence between the upper and lower jaw and the dorsal  and ventral  segments of the gill arch skeleton was recognised over a century ago and has led to various scenarios of jaw evolution by transformation of an anterior member of a hypothetical primitive series of gill-bearing arches e.g. . There iThe notion of an ancestral gill arch-like nature of the vertebrate mandibular arch, however, remains contentious on other grounds. Extant jawless vertebrates (i.e. cyclostomes – lampreys and hagfishes) do not have a gill associated with their mandibular arch derivatives, and in a rare instance of preservation of putative gill impressions in the pharyngeal region of a stem-vertebrate fossil, such impressions are apparently lacking from the anterior-most arch . FurtherUnlike cyclostomes, however, many jawed vertebrates do possess a gill-like epithelial elaboration called a ‘pseudobranch’ in close association with their mandibular arch derivatives. In teleost fishes, the pseudobranch resides in a subopercular cavity , while iThe close resemblance of the pseudobranch to the lamellae of the hyoid and gill arches, its location at the posterior margin of mandibular arch-derived structures and its formation, ancestrally, within a gill slit serial homologue have led to its interpretation as a reduced or vestigial mandibular arch-derived gill . But thePatterns of serial homology may be recognised based on the apparent repetition or correspondence of anatomical features within an organism and are generally attributed to continuity or sharing of underlying developmental information . In manyLeucoraja erinacea. We demonstrate unequivocally, by histological and gene expression analyses and cell lineage tracing, that the skate pseudobranch is mandibular arch derived, and that it shares cell types and gene expression patterns with the gill lamellae of the hyoid and gill arches. We further show that the spiracular cartilage, which supports the pseudobranch in skate, is patterned by a shh-expressing epithelial signalling centre, in a manner that closely parallels the way in which hyoid and gill arch branchial rays are patterned by a shh-expressing gill arch epithelia ridge (GAER). Taken together, our findings strongly support the pseudobranch as a mandibular arch-derived gill serial homologue and, we argue, as a vestige of an ancestral gill arch-like nature of the vertebrate mandibular arch.Here, we have investigated the nature and development of the pseudobranch in a cartilaginous fish, the skate In the skate, the developing pseudobranch is first apparent at S29 as a thickening of the epithelium of the posterior face of the mandibular arch A, and byfoxl2 is expressed in the core mesoderm and presumptive gill-forming epithelium of the pharyngeal arches of cartilaginous fishes (foxl2 expression by mRNA in situ hybridisation (ISH) in skate embryos at S24 reveals these expression patterns not only in the developing hyoid and gill arches, but also in the developing mandibular arch , with epas1/hif2a expression likely reflecting cells that function in hypoxia sensitivity and/or stress response , which function to mediate physiological responses to hypoxia, and which are identifiable by their immunoreactivity for serotonin (5-HT) and synaptic vesicle glycoprotein 2 (SV2) . In skatresponse , and gatid gland .Fig. 4.in situ hybridisation on sections revealed conservation of the above gene expression features in the skate pseudobranch . These observations point to shared cell types and gene expression features – and, by extension, function – between the differentiated pseudobranch and gills of the skate, consistent with the serial homology of these structures.Immunostaining and dobranch F. We foudobranch G-Giii, ashh-expressing epithelial signalling centre called the gill arch epithelial ridge (GAER)  . In thesskeleton .shh expression in the developing mandibular arch of the skate. We found that, at S22, shh was broadly expressed in the posterior epithelium of the skate mandibular arch  mesenchymal cells underneath the mandibular arch GAER , as well as in the connective tissue surrounding the spiracular cartilage. These experiments indicate that, like gill arch branchial rays, the spiracular cartilage develops from GAER shh-responsive mesenchyme of the mandibular arch.To test the fate of mesenchyme underneath the s at S27 A. We labrch GAER B-D, rearshh signalling patterns the spiracular cartilage in a manner that is comparable with the GAER patterning of hyoid and gill arch branchial rays. We have previously shown in skate that from S22 to S27, successively earlier inhibition of hedgehog signalling by cyclopamine treatment resulted in a progressively greater reduction of gill arch mesenchymal cell proliferation, and a corresponding reduction in the number of branchial rays on each arch . Conversely, cyclopamine treatment at S22 and S27 led to striking spiracular cartilage defects (n=3/6) or present as substantially reduced nodules of cartilage (n=3/6), while defects in embryos treated at S27 ranged from complete loss of the spiracular cartilage (n=2/12) to a reduction of the characteristic leaf-like sheet of cartilage, leaving only the slender endpoints connected by a thin bridge of cartilage (n=10/12). In contrast, cyclopamine treatment at S29 resulted in spiracular cartilages with some slightly abnormal morphology  skate embryos showed no observable spiracular cartilage defects A,A′, and defects B-C′, and defects E. After rphology D,D′, butrphology E. These Anatomical and histological similarities between the pseudobranch and gills are striking, and have led to hypotheses of their homology and, by extension, of an ancestral gill-bearing mandibular arch . The latfoxl2-expressing epithelium on the posterior margin of the mandibular arch in skate . However, regardless of this direct evidence of embryonic origin, the vascular and neural characteristics described above need not necessarily preclude a mandibular arch origin of the pseudobranch. In most fishes, blood supply to the afferent pseudobranchial artery does derive from the efferent hyoidean in adults, but this configuration arises from an embryonic pseudobranch vascular condition that more closely resembles that of the gills . In addition, although mandibular arch-derived structures do tend to be innervated by the trigeminal nerve, there are exceptions: e.g. the maxillary barbels of teleosts develop from the mandibular arch but are innervated by the facial nerve , recognisable based on their morphology and co-expression of 5HT and SV2 : shh signalling maintains proliferation of branchial ray progenitors, and progressively earlier inhibition of hedgehog signalling with the small molecule antagonist cyclopamine results in a successively greater reduction of the branchial ray series  mesenchyme, and that progressively earlier inhibition of hedgehog signalling with cyclopamine results in a successively greater reduction of the spiracular cartilage. These mandibular arch signalling interactions and spiracular cartilage phenotypes precisely mirror those of the hyoid and gill arch GAERs and branchial rays, and we therefore argue that the former are clear serial homologues of the latter  eggs were obtained from the Marine Resources Centre of the MBL. Staging was carried out according to in ovo according to All animal work complied with protocols approved by the Institutional Animal Care and Use Committee at the Marine Biological Laboratory . Skate (All embryos for paraffin histology were dehydrated into 100% ethanol (or changed from 100% methanol to 100% ethanol) before clearing in Histosol . Embryos were washed three times for 20 min each in Histosol at room temperature, twice for 30 min each in 1:1 Histosol:paraffin at 60°C and then then moved into molten paraffin wax for overnight at 60°C. The following day, embryos were washed four times for 1 h each in paraffin, and then embedded in peel-a-way moulds. Embedded embryos were sectioned at 8 μm on a Leica RM2125 rotary microtome and sections were mounted on Superfrost Plus slides (Fisher). Histochemical staining was performed using the modified Masson's trichrome staining protocol of in situ hybridisation for foxl2 (MW457610), gata2 (MZ501824), shh (EF100667) and ptc (EF100663) was performed in wholemount as described by in situ hybridisation by chain reaction on paraffin wax-embedded sections was carried out according to Leucoraja erinacea hif2a/epas1 , gata3  and gcm2 , and HCR amplification hairpins were purchased from Molecular Instruments (https://www.molecularinstruments.com). Slides to be used for immunofluorescence were dewaxed in histosol and rehydrated through a descending ethanol series into 1×PBS+0.1% Triton X-100 (PBT). For antigen retrieval, slides were preheated in distilled water for 5 min at 60°C, transferred to prewarmed antigen retrieval solution (10 mM sodium citrate at pH 6.0) and incubated for 25 min at 95°C. Slides were then cooled at −20°C for 30 min. Slides were rinsed three times for 10 min each in PBT, blocked for 30 min in 10% sheep serum and incubated in primary antibody under a parafilm coverslip in a humidified chamber overnight at 4°C. The next day, slides were rinsed three times for 5 min each in PBT and incubated in secondary antibody under a parafilm coverslip in a humidified chamber overnight at 4°C. Slides were then rinsed three times for 10 min each in PBT and coverslipped with Fluoromount G containing DAPI (Southern Biotech). Primary and secondary antibodies were diluted as follows: rabbit anti-5HT ; mouse anti-SV2 ; goat anti-rabbit 488 ; and goat anti-mouse 633 .Chromogenic mRNA In ovo cyclopamine treatment was performed according to in ovo cyclopamine concentration of ∼20 μM, 25 μl of 9 mM stock solution of cyclopamine (LC labs) in dimethylsulfoxide (DMSO) was injected into skate eggs at S22, S27 or S29, using a syringe and 30-gauge needle. For control embryos, an equivalent volume of DMSO was injected into the egg. Cyclopamine-treated embryos were grown to S32, and surviving embryos  were fixed as described above and analysed for skeletal defects by skeletal preparation.For whole-mount staining of the endoskeleton, skate embryos were moved from 100% methanol into 70% ethanol at room temperature. Embryos were then stained with 0.2 mg/ml Alcian Blue 8GX (Sigma) in acetic ethanol  for 24 h with gentle rocking at room temperature. Embryos were destained in acetic ethanol for 24 h, rehydrated through a descending ethanol series, washed in double distilled water, and digested in 1% w/v trypsin in 2% w/v sodium tetraborate for 20 min. Embryos were then cleared in a graded 0.5% w/v KOH:glycerol series  for 24 h/step. Specimens were stored in 80% glycerol at 4°C and manually dissected for photography.n=3) spiracular cartilages at S32 (P=0.305).For the statistical analysis, the dissected left spiracular cartilage of each embryo was imaged on a Leica M165FC stereomicroscope, and the surface area of each cartilage was measured using Fiji  and norm


In the treatment of cervical spondylotic radiculopathy (CSR), spinal endoscopy has been developed vigorously in the past 30 years. However, its effectiveness and subsequent problem of cervical spine stability have always been the controversial hotspots. This study aims to conduct a retrospective study using posterior cervical full-endoscopic technique for the treatment of CSR with foraminal bony stenosis, and evaluate its clinical effect and application value.All 22 patients treated for CSR with foraminal bony stenosis using posterior cervical full-endoscopic technique were analyzed since Dec 1, 2016, to Apr 30, 2020. The data collection included operation time, length of stay, wound healing, surgical complications, visual analog scale (VAS), Japanese Orthopaedic Association (JOA) scores, intervertebral foramen diameter, intervertebral foramen area and cervical instability. The relevant indicators were observed on admission, at postoperative 1 week and 3 months, and at the last follow-up.p < 0.05). There were no statistical differences between VAS or JOA at different postoperative time points (p > 0.05). The height, anteroposterior diameter and area of intervertebral foramen after operation were significantly increased compared with before operation (p < 0.05).The operation time was 141.6 ± 13.7 min. The length of stay was 6.0 ± 2.5 days. VAS and JOA at different time points after operation were decreased compared with before operation (Posterior cervical full-endoscopic technique shows the advantages of smaller invasion, faster recovery, significant effectiveness and fewer complications in our study. Meanwhile, it has little influence on the ROM and stability of the cervical spine. Therefore, it is a minimally invasive, safe and effective surgical method for the treatment of CSR with foraminal bony stenosis. Cervical spondylotic radiculopathy (CSR) is mainly characterized by axial pain and/or radicular pain, which is caused by the compression of nerve roots due to cervical disc herniation or cervical spinal stenosis. The incidence rate of cervical spondylosis is about 3.8%–17.6%, of which CSR accounts for 60% of all types of cervical spondylosis . AnterioTherefore, our study aims to conduct a retrospective study using posterior cervical full-endoscopic technique for the treatment of CSR with foraminal bony stenosis, and evaluate its clinical effect and application value.We obtained ethical approval exemption from our ethics committee to perform this study since we didn't have direct contact with the participants. Patients treated for CSR in database records of our hospital were retrospectively analyzed from Dec 1, 2016, to Apr 30, 2020.Inclusion criteria were as follows: (1) the diagnostic criteria referred to the CSR clinical guideline of NASS Evidence-Based Guideline Development Committee ; (2) ageThe posture refers to professor Ruetten's method . The patThe focus is located by anteroposterior and lateral radiograph. An 8 mm incision is made after local anesthesia. Place in and position the primary dilator accurately, then screw in the working sleeve. Subsequently, insert the endoscope. After connecting the lavage device, use nucleus pulposus forceps and a high-radiofrequency electrotome to clean the muscle and soft tissue on the surface of lamina. Once the “V” point is clearly identified, use a high-speed drill to expand the lamina space from head to tail. Generally, grind off the lower edge of upper lamina and the upper edge of lower lamina by about 3 mm respectively. Grind off the inner edge of lateral mass by 3 mm to the outside simultaneously. Remove the ligamentum flavum with basket forceps and gun forceps. Use the nerve hook to separate the scar on nerve and identify the spinal cord and exiting root. Next, grind off the upper edge of the lower cervical pedicle to create a potential channel from the axilla of nerve root to the rear of the intervertebral space. Gently rotate the working sleeve to protect the spinal cord and nerve root, adjust the lens to aim at the intervertebral space, excise the herniated nucleus pulposus and annulus fibrosus with nucleus pulposus forceps, and cut off the posterior osteophyte of the intervertebral space with an osteotome or a drill. Then explore again whether there is residual nucleus pulposus or osteophyte around the nerve root, perform annulus fibroplasty with the radiofrequency electrotome, and inject dexamethasone after hemostasis. Finally, remove the working sleeve followed by wound closure .Routine swelling and pain relief and neurotrophic treatment are performed after the operation. Patients can begin out-of-bed activities with soft neck braces on the 2nd postoperative day. The neck brace should be worn for 4 weeks. The postoperative outpatient and telephone follow-up last for 6–18 months, with an average of 11.2 ± 6.5 months.Operation time and length of stay were recorded. Relevant surgical complications such as infection, spinal cord and nerve injury, and systemic manifestations were also collected. The pain was evaluated based on visual analog scale (VAS). In addition, Japanese Orthopaedic Association (JOA) scores was used to assess the improvement in the motor, sensory and bladder dysfunction of patients. VAS and JOA score were recorded routinely on admission, at postoperative 1 week and 3 months, and at the last follow-up.via, software version:VB40, Siemens Healthineers, Forchhiem, Germany), performing multiplanar reconstruction (MPR) in the vertical direction of the intervertebral foramen, manually found and determined the best display layer. Then we used the area measuring tool to sketch along the contour curve of the intervertebral foramen, and obtained the contour area (cm2) after drawing. Finally, we recorded the results.The intervertebral foramen height and anteroposterior diameter, and intervertebral foramen area were measured pre- and postoperatively (1 week). The sagittal section of the internal edge of the pedicle on operated side was defined as the measuring plane, and the height and diameter were measured based on this plane. For intervertebral foramen area, we transferring the thin-layer scanning DICOM data to the workstation  was used to judge whether there was postoperative segmental instability .χ2 test, Fisher's exact probability method, or non-parametric testing. Differences in variables before and after the operation were analyzed through one-way ANOVA or paired t-test. Meanwhile, p < 0.05 was considered statistically significant.Statistical Packages of Social Sciences (SPSS) software (version 22.0) was used to analyze the collected data. Measurement data were tested for normality of distribution. Data with normal distribution were expressed as mean ± standard deviation (`x ± s). Counting data were described as percentages or constituent ratios, and were analyzed using the A total of 22 consecutive cases from our database of patients were involved in the current study, including 15 males and 7 females, with an average age of 49.6 ± 9.2 years (ranged from 36 to 78 years). All cases were single-level unilateral CSR with foraminal bony stenosis, including 1 case of C3/4 level, 4 cases of C4/5 level, 15 cases of C5/6 level and 2 cases of C6/7 level .The radiating pain and neurological symptoms of all cases were significantly improved after operation. There were no serious complications such as spinal cord and nerve injury. The operation time was 141.6 ± 13.7 min and the length of stay was 6.0 ± 2.5 days. All wounds healed by first intention without postoperative infection. One case suffered a transient burning sensation of the affected extremity after operation, and the pain was not significantly relieved compared with that before operation. CT and MRI showed that the decompression was sufficient, but there was a low signal area in the intervertebral foramen on MRI. The patient was given fluid gelatin to stop bleeding during operation, and it was considered that the excessive fluid gelatin and insufficient lavage resulted in local hematoma. Intravenous dexamethasone, dehydration and analgesic treatment were given postoperatively. The patient's pain was significantly improved on the 3rd day after operation, and all pathological symptoms had disappeared at the postoperative 3-month follow-up .p < 0.05), which indicated significant alleviation of pain and improvement of motor function. There were no statistical differences between VAS scores or JOA at different postoperative time points (p > 0.05) (VAS scores and JOA at different time points after operation were decreased compared with before operation ( > 0.05) .p < 0.05). The postoperative intervertebral foramen area was significantly larger than that before operation (p < 0.05), which clearly indicated that the spinal canal and intervertebral foramen are fully decompressed under endoscope  smaller invasion, less injury to the facet joints and posterior soft tissue, and better preservation of the stability of the cervical spine; (2) the shorter length of stay and the earlier functional training, which can not only enhance recovery after surgery, but also save medical resources; (3) preserving the motion segment, which greatly reduces the influence on ROM of the cervical spine and the degeneration of adjacent segments; (4) The magnification effect of endoscopic visual field and continuous hydraulic pressure can effectively maintain the clear view of surgery, in order to increase the identification of tissue structure and reduce the risk of nerve and vascular injury; (5) the 360° intervertebral foraminotomy can achieve fully decompression under minor interference of the nerve root.Admittedly, there are some deficiencies in the posterior cervical full-endoscopic technique, especially the long learning curve. The technique places enormous demands on operation under endoscope and use of grinding drill. Those with insufficient experience performing the surgery blindly will cause common complications including incomplete decompression, nerve root irritation, infection, etc., or even disastrous consequences such as spinal cord injury and quadriplegia , 19. In The current study has several limitations and it could be altered in some ways to better ascertain the conclusion. First of all is its relatively small sample size. We limited the indications to foraminal bony stenosis in order to treat CSR more accurately. However, appropriate extension of indications and larger sample population will allow for more meaningful statistical testing and smaller deviation. Another limitation is the short follow-up period. The longest period in this study was 18 months since we initially believed that longer period of follow-up would reduce patients' compliance and make data collection more difficult. Nevertheless, longer period up to 2 or 3 years may improve the reliability of evidence and be more likely to find the occurrence of adjacent segment disease, so as to confirm the long-term effect of this method. Therefore, large sample, multicenter and long follow-up studies should be performed in our future clinical work to evaluate its clinical application value.Posterior cervical full-endoscopic technique shows the advantages of smaller invasion, faster recovery, significant effectiveness and fewer complications in our study. Meanwhile, it has little influence on the ROM and stability of the cervical spine. Therefore, it is a minimally invasive, safe and effective surgical method for the treatment of CSR with foraminal bony stenosis. However, more improved studies should be conducted to provide orthopedic surgeons with the best evidence-based information.