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OBJECTIVE@#To establish an intestinal organoid model that mimic acute graft versus host disease (aGVHD) caused intestinal injuries by using aGVHD murine model serum and organoid culture system, and explore the changes of aGVHD intestine in vitro by advantage of organoid technology.@*METHODS@#20-22 g female C57BL/6 mice and 20-22 g female BALB/c mice were used as donors and recipients for bone marrow transplantation, respectively. Within 4-6 h after receiving a lethal dose (8.0 Gy) of γ ray total body irradiation, a total of 0.25 ml of murine derived bone marrow cells (1×107/mice, n=20) and spleen nucleated cells (5×106/mice, n=20) was infused to establish a mouse model of aGVHD (n=20). The aGVHD mice were anesthetized at the 7th day after transplantation, and the veinal blood was harvested by removing the eyeballs, and the serum was collected by centrifugation. The small intestinal crypts of healthy C57BL/6 mice were harvested and cultivated in 3D culture system that maintaining the growth and proliferation of intestinal stem cells in vitro. In our experiment, 5%, 10%, 20% proportions of aGVHD serum were respectively added into the organoid culture system for 3 days. The formation of small intestinal organoids were observed under an inverted microscope and the morphological characteristics of intestinal organoids in each groups were analyzed. For further evaluation, the aGVHD intestinal organoids were harvested and their pathological changes were observed. Combined with HE staining, intestinal organ morphology evaluation was performed. Combined with Alcian Blue staining, the secretion function of aGVHD intestinal organoids was observed. The distribution and changes of Lgr5+ and Clu+ intestinal stem cells in intestinal organoids were analyzed under the conditions of 5%, 10% and 20% serum concentrations by immunohistochemical stainings.@*RESULTS@#The results of HE staining showed that the integrity of intestinal organoids in the 5% concentration serum group was better than that in the 10% and 20% groups. The 5% concentration serum group showed the highest number of organoids, the highest germination rate and the lowest pathological score among experimental groups, while the 20% group exhibited severe morphological destruction and almost no germination was observed, and the pathological score was the highest among all groups(t=3.668, 4.334,5.309,P<0.05). The results of Alican blue staining showed that the secretion function of intestinal organoids in serum culture of aGVHD in the 20% group was weaker than that of the 5% group and 10% of the organoids, and there was almost no goblet cells, and mucus was stainned in the 20% aGVHD serum group. The immunohistochemical results showed that the number of Lgr5+ cells of intestinal organoids in the 5% group was more than that of the intestinal organoids in the 10% aGVHD serum group and 20% aGVHD serum group. Almost no Clu+ cells were observed in the 5% group. The Lgr5+ cells in the 20% group were seriously injuried and can not be observed. The proportion of Clu+ cells in the 20% group significantly increased.@*CONCLUSION@#The concentration of aGVHD serum in the culture system can affect the number and secretion function of intestinal organoids as well as the number of intestinal stem cells in organoids. The higher the serum concentration, the greater the risk of organoid injury, which reveal the characteristics of the formation and functional change of aGVHD intestinal organoids, and provide a novel tool for the study of intestinal injury in aGVHD.
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Mice , Female , Animals , Mice, Inbred C57BL , Bone Marrow Transplantation , Graft vs Host Disease , Stem Cells , OrganoidsABSTRACT
Objective:To investigate the application and surgical effect of lobed anterolateral femoral chimeric perforator flap in repairing the complex soft tissue defects of limbs.Methods:Clinical materials of 52 cases with complex soft tissue defects of limbs were retrospectively reviewed from October 2013 to October 2020. Thirty-two males and 20 females were reviewed, and the age of them ranged from 6 to 65(mean 39) years old. The injuries include 17 cases of traffic accidents, 15 machine crush injuries, 12 engineering accidents, and 8 extensive resection of malignant tumors. The size of soft tissue defects ranged from 26.0 cm×10.0 cm to 35.0 cm×15.0 cm. Lobed anterolateral thigh chimeric perforator flaps or lobed Flow-through chimeric perforator flaps were designed according to the soft tissue defects and blood circulation of the distal extremities. Start from 8-month post-operation, upper limb function was assessed according to the Upper Limb Function Assessment Standard of Hand Surgery Branch of Chinese Medical Association and Disability of arm shoulder and hand(DASH) score scale, and Fugl-Meyer score system was used to evaluate the lower limb function. Long-term follow-up was conducted through WeChat video call and outpatient clinic evaluation.Results:All flaps survived. One patient with crush injury got delayed wound healing due to deep hemaetoma after surgery, and the rest were healed at the first stage. All the cases entered followed-up, ranged from 6 to 20(mean 8) months. The appearance of the flaps was satisfactory, and the color of the flaps has no significant difference compared with the recipient area. No recurrence of tumor was reported during the follow-up period. According to the Upper Limb Function Assessment Standard of Hand Surgery Branch of Chinese Medical Association, there were 21 excellent cases and 8 good cases, and the total excellent and good rate was 100%. The DASH score of upper limbs ranges from 11.5 to 45.1(mean 25.4±13.7), and the Fugl-Meyer score of lower limbs range from 18.6 to 31.8(mean 26.2±11.2) at the last follow-up.Conclusion:The complex soft tissue defect of limbs is a common complication caused by high-energy trauma, which is difficult to repair. The lobed anterolateral thigh chimeric perforator flap through a Decompose-Combine and arrange method for reconstruction, which is flexible in flap design. A variety of tissue flaps can be obtained by anastomosing only one group of blood vessels. It is an ideal choice for repairment of complex soft tissue defects of limbs.
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Objective:To explore the significance of multidisciplinary team for the diagnosis and treatment of congenital band syndrome.Methods:Five children with congenital circular band syndrome admitted in the First Affiliated Hospital of the Chinese People′s Liberation Army Air Force Medical University from January 2017 to January 2020 were retrospectively analyzed.They were diagnosed, treated and followed up by a multidisciplinary team including the department of obstetrics and gynecology, ultrasound, and orthopedics from the fetal stage.Results:One case was found with gradually aggravated ring band that affects the blood circulation of distal limbs during fetal examination, who was promptly performed with partial band release under fetoscopy.All cases had Patterson type Ⅰ ring bands at birth, and distal limbs did not have obvious deformity.They were performed with selective annular band resection, and postoperatively followed up for an average of 10.6 (8-13) months.The annular depression of the skin disappeared, and the appearance and function recovered satisfactory.According to the monitoring of side effects scale (Moses), 4 cases were excellent, 1 case was good, and the excellent and good rate was 100%.Conclusions:The multidisciplinary team for the diagnosis and treatment of congenital band syndrome can maximize the professional advantages of physicians with diffe-rent specialties, make early diagnosis and treatment, minimize the compression of the ring band on the limbs, and avoid serious limb deformities.It is worthy of clinical application.
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Objective::To explore the mechanism of Suanzaoren Tang in improving learning and memory impairment induced by sleep deprivation based on Toll-like receptor (TLR4)/nuclear transcription factor-κB (NF-κB) signaling pathway. Method::The experimental rats were randomly divided into blank group, model group, melatonin group (2.5×10-4 g·kg-1·d-1) and Suanzaoren Tang group (12.96 g·kg-1·d-1). Except the blank group, the chronic sleep deprivation model was established in other groups. After 28 days of continuous administration, the learning and memory of the rats were assessed by Morris water maze. The expressions of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α)in serum were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expression levels of TLR4, NF-κB p65 and nuclear transcription factor inhibitory protein α (IκBα) in the hippocampus of rats were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expressions of TLR4, IκBα, p-IκBα and NF-κB p65 in the hippocampus of rats were detected by immunohistochemistry (IHC). Result::Compared with the blank group, the platform latency, total swimming distance and the first landing time of the model group increased significantly (P<0.01), while the number of crossing platforms and target quadrant time decreased significantly (P<0.01). Compared with the model group, the platform latency, total swimming distance and the first platform time were reduced in the melatonin group and the Suanzaoren group (P<0.05, P<0.01), while the number of crossing platforms and target quadrants time were increased(P<0.05, P<0.01). Compared with the blank group, the expression levels of TNF-α, IL-1β and IL-6 in the model group were significantly increased (P<0.01). Compared with the model group, the levels of TNF-α, IL-1β and IL-6 in the serum of melatonin group and Suanzaoren Tang group decreased (P<0.05, P<0.01). Compared with the blank group, mRNA and protein expression levels of TLR4 and NF-κB p65 in the hippocampus of the model group were increased (P<0.01), while mRNA and protein expression levels of IκBα were decreased (P<0.01). Compared with the model group, the mRNA and protein expressions of TLR4 and NF-κB p65 in the hippocampus of melatonin group and Suanzaoren Tang group decreased (P<0.05, P<0.01), while the mRNA and protein expression levels of IκBα increased (P<0.05, P<0.01). Compared with the blank group, the protein expression level of p-IκBα in the hippocampus of the model group was significantly increased (P<0.01). Compared with the model group, the protein expression level of IκBα in the hippocampus of melatonin group and Suanzaoren Tang group was increased (P<0.05, P<0.01). Conclusion::Suanzaoren Tang can improve learning and memory impairment induced by sleep deprivation in rats, and its mechanism may be related to its inhibition of TLR4/NF-κB signaling pathway in hippocampus.
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Objective:To compair the outcomes of repairing soft tissue defects of hand between a biodegrad- able repair patch—porcine small intestinal submucosa (SIS) and skin grafting.Methods:From December, 2017 to December, 2018, 36 cases of hand soft tissue defect were treated and analyzed retrospectively. According to the defect area and treatment methods, 36 cases were divided into 2 groups: SIS group (21 cases) and grafting group(15 cases). In SIS group, the area of soft tissue defects was 2.0 cm×1.5 cm-9.0 cm×3.5 cm with an average of 5.3 cm×2.1 cm, treat- ed with SIS; In grafting group, the area of soft tissue defects was 9.0 cm×4.0 cm-16.0 cm×9.0 cm with an average of 12.0 cm×8.5 cm, treated with autologous skin grafting after wet dressing. Wound healing was evaluated at 14, 21 and 28 days, and 3 months after the surgery according to the appearance of colour, elasticity, sensory recovery and prog- noses of partial tendon exposure.Results:All patients were followed-up for 3 to 10 months, with an average of 5 months. All wounds in both groups were completely healed; the appearance was normal, and the skin elasticity and sensation had recovered. Sensation recovery in SIS group: 14 cases were good (66.6%), 5 cases were fine (23.8%), and 2 cases were bad (9.6%); in grafting group: 9 cases were good (60.0%), 4 cases were fine (26.0%), and 2 cases were bad (14.0% ). Wound healing effect in SIS group: 14 cases were good, 5 cases were fine, and 2 cases were bad; in grafting group: 9 cases were good, 4 cases were fine, and 2 cases were bad.Conclusion:The SIS patch can be used in the reconstruction of soft tissue defects in hand. There was no significant difference in colour compared to the sur- rounding skin and left no scar. The patch is an ideal repair material for superficial skin defects.
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Objective To evaluate the clinical safety of the collagen nerve scaffold with longitudinally oriented microchannels in bridging peripheral nerve defect.Methods Five patients with 8 peripheral nerve defects of 18 to 30 (mean,23.8) mm in length were involved in the pilot study and treated from July,2017 to March,2018,including 6 digital nerves and 2 medial antebrachial cutaneous nerves.The defects were repaired with the collagen nerve scaffold with longitudinally oriented microchannels independently developed.Routine therapy of prophylactic systemic antibiotics but no immunosuppressive drugs was given to all patients post-operatively.All patients were followed-up by regular review in the outpatient department combined with WeChat and telephone.The clinical safety of the nerve scaffold was preliminarily evaluated through observing the condition of the healing of the local wound and the whole body.The blood routine examineation and biochemical test were detected.The statistical analysis of the measurement data was performed by the analysis of variance of repeated measurement data,and the difference was statistically significant when P＜0.05.Results All patients were followed-up for 7 to 15 months (average,10 months).No adverse events such as infection,allergy,damage of liver and kidney function occurred.The operative incisions healed primarily,with no redness,exudation and rupture in the local area.There was no systemic symptoms such as fever,nausea,vomiting,skin itching,etc.The results of blood routine tests and biochemical tests were normal.The data of tests was compared,and the difference was not statistically significant (P＞0.05).Conclusion The preliminary study shows that it is clinically safe to bridge peripheral nerve defects with the collagen nerve scaffold with longitudinally oriented microchannels.
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Objective To investigate the feasibility and technique of using the anterolateral thigh perforator flap pedicled with the lateral branch of the lateral circumflex femoral artery (LBLCFA) to repair soft tissue defect of extremities.Methods Eighty-six cases of anterolateral thigh perforator flap transplantation were performed from May,2014 to May,2018.A total of 37 cases of soft tissue defect of extremities were treated by anterolateral thigh perforator flap used the LBLCFA as vascular pedicle,of which there were 19 cases of upper limbs and 18 of lower limbs.There were 27 cases of defects caused by trauma,and 10 by soft tissue tumor resection.The flaps were designed centering around the point proximal to the midpoint of the iliac-patellar line.The dimensions of soft tissue defect were from 9.0 cm×6.0 cm to 26.0 cm×10.0 cm,and the flap were from 10.0 cm×7.0 cm to 27.0 cm×11.0 cm.The length of vascular pedicle ranged of 7.0-13.0 cm,with an average of 11.5 cm.The donor sites were directly sutured.All of the patients were followed-up regularly in the outpatient department.Results All the flaps survived and the donor sites were primarily healed.Of these 37 cases,2 trauma patients and 2 patients treated with local radiotherapy had poor wound healing,but still healed after multiple dressing changes.All the patients were followed-up for 3-26 months,with an average of 13 months.The texture,color and elasticity of the flap were similar to the surrounding tissue of the recipient sites,while only a linear scar remained at the donor sites.Ten tumor patients were treated with routine radiotherapy and chemotherapy after the repairation;there was no tumor recurrence during the follow-up period.Conclusion The LBLCFA gives off a relatively large and thick perforator proximal 5.0-7.0 cm of the iliacpatellar line,which locates in the upper lateral side,travels parallel to the vastus lateralis,and give off the skin and muscular perforators.The lateral branch can be used as pedicle to make into perforator flap or chimeric flap,which provides a novel selection of vascular pedicel for anterolateral thigh perforator flap.When the descending branch does not provide a thick and large perforator,the LBLCFA has important practical value and is worth utilizing in the clinic.
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Objective To explore the influence of ROI size on the diagnostic performance of shear wave elstography (SWE) of breast masses.Methods A total of 225 consecutive patients with 239 solid breast masses underwent SWE before surgery,and then quantitative SWE parameters,including the maximum stiffness (Emax),mean stiffness (Emean),ratio of stiffness of the mass to stiffness of surrounding fat (Eratio) and standard deviation (SD),were measured by two methods,one method was using larger ROI to include the largest part of a mass,and the other was placing fixed ROI=2 mm over the stiffest part of the lesion to measure.The diagnostic performance of the two methods were evaluated by ROC curve,and the cut-off value,sensitivity and specificity of each SWE parameter were obtained.Results There were no statistically significant differences between the performance of Emax,Emean and SD from the two different methods (all P＞ 0.05),the performance of Eratio of the two methods showed significant difference (P=0.046).The significantly higher specificity of Emax and Emean in the larger ROI were found,and the significantly higher sensitivity of SD and Eratio in the ROI=2 mm was found.Furthermore,the diagnostic performance of combination of SWE and BI-RADS were not greatly affected by the ROI size,the difference had no statistical significance (all P＞0.05).Conclusion The ROI size only affects the diagnostic performance of Eratio,and donot affect the diagnostic performance of the quantitative SWE parameters (Emax,Emean,SD) and combination of SWE and BI-RADS.
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Objective To investigate the preventive effect of the artificial nerve canal on irritating neuralgia after neurilemmoma resection.Methods Thirty patients diagnosed as neurilemmoma were divided into artificial nerve group and control group at randomly.There were 15 patients in each group.The nerves in artificial nerve group were enclosed using the artificial nerve besides neurilemmoma resection.The nerves in control group only had neurilemmoma resection.All patients were assessed with VAS for the degree of irritating neuralgia at 3 time points after operation.Calculate the incidence of irritating neuralgia and analyze the data using statistical methods.Results In control group,the VAS scores were 7.5,5.1,2.8,and the incidences of irritating neuralgia were 80％,67％,40％,respectively.In artificial nerve group,the VAS scores were 3.2,1.5,0,and the incidences of irritating neuralgia were 33％,13％,0.The VAS scores and the incidences of irritating neuralgia of two groups had significant difference by statistical analysis (P ＜ 0.05).Conclusion The artificial nerve canal can relieve the degree and decrease the incidence of irritating neuralgia after neurilemmoma resection,which is a useful method to prevent irritating neuralgia in clinic.
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Objective To evaluate the application of high-resolution ultrasonography in the diagnosis of nerve root lesion of cervical vertebra intervertebral foramen part. Methods Twenty-seven patients with root avulsion injury of brachial plexus intervertebral part and 8 patients with schwannoma of brachial plexus root were examined with high-resolution ultrasonography. Thirty healthy volunteers were involved for the sonographic imagings of normal brachial plexus intervertebral part. The ultrasound findings of the patients before operation were compared with surgical outcomes. Results The normal intervertebral part of brachial plexus were shown as hypoechoic tubular strucuture, which could be identified with cervical artery and transverse process used as landmarks for the intervertebral part of brachial plexus root. The detection ratios of normal intervertebral part for C5-C8 and outside part of braehial plexus were 100％. In 27 patients with root avulsion injury of intervertebral brachial plexus, the intervertebral nerve root were enlarged as hypoechoic area, with nerve dissected partly or disappearing, distal nerve parts outside the intervertebral foramen swelling, edema,or cerebrospinal fluid assembling as cystic masses. In 8 patients with schwannoma of brachial plexus root part, the spinal ganglia outside intervertebral foramen were shown as tumor-like hypoechoic mass, with blood supply shown inside. Conclusions High-resolution ultrasonography is of great value for early and convenient diagnosis of root avulsion injury and mass of brachial plexus intervertebral part, especially in community hospitals,which could provide important information for clinical therapy.
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Objective To analyze sonographic imagings of traumatic neuromas of peripheral nerves,and to investigate the applied values of uhrasonography in the traumatic neuroma of peripheral nerve. Methods The sonographic imagings of 38 patients who had evidences of traumatic neuromas in peripheral nerves were retrospectively reviewed by the last diagnostic results of operations. Results Among 38 cases, 2 incomplete traumatic neuromas were not detected, and 1 incomplete traumatic neuroma was misdiagnosised as neurilemoma by ultrasonography. The coincidence of uitrasonography in incomplete traumatic neuroma and complete severed neuroma was 82.4% and 100%. With regard to the echogenicity of 38 traumatic neuromas of peripheral nerves, the sonographic characteristics of neuromas were all hypoechoic. Thirty-three neuromas appearanced like shuttles and 5 neuromas appearaneed like humps. Conclusions The traumatic neuroma has distinctive ultrasonic appearance. Ultrasonography is useful to direct the diagnosis and treatment of traumatic neuroma.
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Objective To explore a new method of functional reconstruction of hand digits and joints with free transfer of foot tissues so as to increase the success rate of the operation.Methods After micro-anatomic study of the plantar and dorsal metatarsal arteries,retrograde and free grafts of foot tissues pedicled with plantar metatarsal arteries were designed and applied in transplantation to treat 76 cases of hand digital or joint defects.The surgeries included 58 cases of transfer of the second toe,four cases of transfer of composite tissues of the second toe, eight cases of transfer of proximal interphalangeal joint,and six cases of nail flap transfer.Results The mi- cro-anatomic study found that the first plantar metatarsal artery was anatomically constant and the diameter of its branch to the second toe was larger than that of the first dorsal metatarsal artery.Flaps survived in 75 of the 76 patients(98.7%),with fine appearance and significantly improved function.One patient who had received free transfer of the second toe to reconstruct the thumb function had to undergo a second repair with infraclavicula skin tube because of refractory arteriospasm of anastomosed vessels.Conclusion Transfer with free retrograde grafts of foot tissues pedicled with plantar metatarsal artery to reconstruct hand functions can effectively improve the success rate of the operation,because it is free of the shortcomings of great anatomic variation of blood vessels and time-consuming and complex procedures in conventional transfer.