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1.
Toxicol In Vitro ; : 105292, 2021 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-34871754

RESUMO

Antibody-drug conjugates (ADCs) are a promising modality for cancers, but the interaction between them and proton pump inhibitors (PPIs), the common adjuvant drugs for cancer treatment, has not been understood. Here, the interactions between PPIs and RC48ADC, a novel HER2-targeting ADC, were quantified in vitro. CCK-8 assay showed that RC48ADC displayed a significant inhibitory effect on the proliferation of SK-BR-3, NCI-N87 and SK-OV-3 cells with the IC50 values of 4.91 ±â€¯1.15 ng/mL, 14.54 ±â€¯0.85 ng/mL and 11.28 ±â€¯0.68 ng/mL respectively. PPIs alone had no significant anti-tumor effect in the dose range of 1.37-1000 ng/mL. When used together, PPIs inhibited the anti-tumor activity of RC48ADC in a dose-dependent manner. And 1000 ng/mL (~Cmax) PPIs significantly recovered RC48ADC-inhibited cell proliferation by (32.85 ±â€¯2.81) % (p < 0.05). However, cimetidine, a non-PPIs gastric acid secretion inhibitor, had no significant inhibitory effect on RC48ADC. Furthermore, omeprazole, rather than cimetidine, significantly reduced the activity of vacuolar H+-ATPase and Cathepsin B compared with the control cells. These results, if confirmed in vivo, indicate that PPIs are antagonists of RC48ADC, even all ADCs, appearing to be due to inhibition of vacuolar H+-ATPase activity. Moreover, cimetidine combined with ADCs instead of PPIs can prevent an adverse drug interaction.

2.
Gels ; 7(4)2021 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-34940307

RESUMO

Three-dimensional cell-laden tissue engineering has become an extensive research direction. This study aimed to evaluate whether chondrocyte spheroids (chondro-spheroids) prepared using the hanging-drop method could develop better cell proliferation and morphology maintenance characteristics, and be optimized as a micro unit for cartilage tissue engineering. Chondro-spheroids were loaded into a cross-linkable hybrid hydrogel of gelatin methacrylate (GelMA) and hyaluronic acid methacrylate (HAMA) in vivo and in vitro. Cell proliferation, aggregation, cell morphology maintenance as well as cartilage-related gene expression and matrix secretion in vitro and in vivo were evaluated. The results indicated that compared with chondrocyte-laden hydrogel, chondro-spheroid-laden hydrogel enhanced proliferation, had better phenotype maintenance, and a more natural morphological structure, which made it appropriate for use as a micro unit in cartilage tissue engineering.

3.
Int Wound J ; 2021 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-34935287

RESUMO

To investigate the clinical application value of different flap transfer and repair techniques in adult patients with chronic osteomyelitis of limbs complicated with soft tissue defects. According to the characteristics and defects of 21 cases, different plastic surgery was applied, including debridement, negative pressure device, and tissue flap to cover wound. Among 21 cases of chronic osteomyelitis complicated with local soft tissue defect, 15 patients were repaired with sural neurotrophic musculocutaneous flap transfer, 2 patients were repaired with medial plantar skin flap transfer, 2 patients were repaired with ilioinguinal skin flap transfer, 1 patient was repaired with z-forming wound, and 1 patient was repaired with soleus muscle flap combined with full-thickness skin graft. All the 21 patients underwent bone cement implantation after dead bone osteotomy. Among them, 19 patients underwent bone cement replacement with 3D prosthesis within 6 months to 1 year after surgery, and 2 patients carried bone cement for a long time. Early intervention, thorough debridement, removal of necrotic or infection, and then selecting the appropriate wound skin flap coverage are important means of guarantee slow osteomyelitis wound healing and for providing a possible way to permanent prosthesis implantation subsequently.

4.
J Phys Ther Sci ; 33(11): 862-869, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34776624

RESUMO

[Purpose] This study aimed to explore the application value of yoga intervention in early postpartum recovery of the levator ani muscle hiatus (LAH) area. [Participants and Methods] Females in natural labor from May 2020 to November 2020 in the Third People's Hospital of Sun Yat-sen University Ultrasound Research Center were prospectively included for a pelvic ultrasound examination. The control group received no intervention. The experimental group received 60-min yoga once a week from week 1 to week 12 postpartum. A pelvic ultrasound examination was performed on the week 6 and week 12 postpartum. The LAH area was measured at rest, during contraction and Valsalva maneuver. [Results] A total of 128 participants who met the inclusion criteria were selected and randomly assigned to the control group (n=66) and the experimental group (n=62) in pre and post intervention design. No significant differences were found in age, parity, body mass index, and fetal weight between the control and experimental groups. Further, no significant difference was observed in the LAH area between the control and experimental groups at rest, during contraction and Valsalva maneuver on the week 6 postpartum. However, the LAH area in experimental group significantly reduced at rest, during contraction and Valsalva maneuver on the week 12 postpartum. The differences of LAH area (date week 6 minus date week 12) in the control group at rest, during contraction and Valsalva maneuver were 0.12 ± 3.12 cm2, 0.80 ± 2.29 cm2, and 0.80 ± 4.22 cm2, while in the control these were 1.95 ± 3.41 cm2, 1.39 ± 1.91 cm2, and 3.81 ± 5.49 cm2, respectively. Compared with control group, the differences of LAH area significantly increased in experimental group at rest and during Valsalva maneuver. [Conclusion] Yoga intervention can help in the recovery of LAH.

5.
Tissue Eng Regen Med ; 2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34618337

RESUMO

BACKGROUND: As a contour-supporting material, the cartilage has a significant application value in plastic surgery. Since the development of hydrogel scaffolds with sufficient biomechanical strength and high biocompatibility, cell-laden hydrogels have been widely studied for application in cartilage bioengineering. This systematic review summarizes the latest research on engineered cartilage constructed using cell-laden hydrogel scaffolds in plastic surgery. METHODS: A systematic review was performed by searching the PubMed and Web of Science databases using selected keywords and Medical Subject Headings search terms. RESULTS: Forty-two studies were identified based on the search criteria. After full-text screening for inclusion and exclusion criteria, 18 studies were included. Data collected from each study included culturing form, seed cell types and sources, concentration of cells and gels, scaffold materials and bio-printing structures, and biomechanical properties of cartilage constructs. These cell-laden hydrogel scaffolds were reported to show some feasibility of cartilage engineering, including better cell proliferation, enhanced deposition of glycosaminoglycans and collagen type II in the extracellular matrix, and better biomechanical properties close to the natural state. CONCLUSION: Cell-laden hydrogels have been widely used in cartilage bioengineering research. Through 3-dimensional (3D) printing, the cell-laden hydrogel can form a bionic contour structure. Extracellular matrix expression was observed in vivo and in vitro, and the elastic modulus was reported to be similar to that of natural cartilage. The future direction of cartilage tissue engineering in plastic surgery involves the use of novel hydrogel materials and more advanced 3D printing technology combined with biochemistry and biomechanical stimulation.

6.
Int Wound J ; 2021 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-34636163

RESUMO

We aimed to explore the clinical significance of the secondary pedicle amputation of the repair of distal defects with pedicled axial flap. Five patients who underwent pedicled axial flap transfer to repair a large area of skin and soft tissue defects in our hospital were included in this retrospective study. Detailed information including general data and clinical data, such as preoperative complication, type of primary wound, the distance between the primary wound and the donor site (cm), postoperative complications, and types of axial flap were collected. The patients had good joint movement at 6 months after pedicle amputation. At 48 hours after transplantation, except for the last patient (NO.5), there were no obvious complications such as blood supply disorder, infection, and incision dehiscence of the patients, and the flaps survived well. Just after pedicle amputation, 3 and 6 months after pedicle amputation, the flaps survived well with good local morphology. Forty-eight hours after operation, part of the distal flap in the last patient (NO.5) was necrotic. After 6 months of pedicle amputation, part of the flap was transferred to the distal wound again. At 6 months after pedicle amputation, these patients could accept local scars even though the scar of the last patient was obvious. The secondary pedicle amputation of the repair of distal defects with axial flap could avoid the compression of the vascular pedicle in the subcutaneous tunnel between the donor site and the primary wound, which may ensure the bold supply and increase the survival rate of the flap.

7.
J Cosmet Dermatol ; 2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-34716978

RESUMO

BACKGROUND: Wound healing problem is one of the main complications after correction of chest wall deformity. Orthopedic flap tissue repair technique has a clear significance in non-healing wound and defect wound and provides a new choice for poor wound healing after orthopedic surgery of chest wall deformity. OBJECTIVE: To investigate, the application value of modified local rotary flap and latissimus dorsi myocutcutaneous flap in the treatment of poor wound healing after orthodontic treatment of chest wall deformity. METHOD: A retrospective analysis was performed on patients who admitted to our department from August 2012 to November 2019 due to non-healing incision after surgery for thoracic deformity. Skin flap was selected according to the size of the wound surface, and the effect of skin flap repair was observed. The clinical data of the included patients were recorded, and the preoperative and postoperative wound conditions were evaluated. RESULTS: This study included 13 patients with chest wall deformity who received plastic surgery tissue using flap technique for wound repair, 11 cases used modified local rotation skin flap, and 2 cases used modified latissimus dorsi myocutaneous flap. The mean age of the 13 patients was 18.54 ± 4.14 years old, the mean body mass index (BMI) was 17.02 ± 2.16 kg/m2 , and the mean preoperative nonunion time of the incision was 64.77 ± 93.01 days. Five patients had positive bacteria culture on the wound surface, including 3 cases of Staphylococcus aureus, 1 case of Pseudomonas aeruginosa, and 1 case of Staphylococcus epidermidis. All the 13 patients achieved primary grade A healing. CONCLUSION: The modified local rotary flap and latissimus dorsi musculocutaneous flap have a significant effect on the postoperative correction of chest wall deformity, which can ensure wound healing while retaining the orthopedic plate to the maximum extent to ensure the effect of the correction.

8.
Int J Biol Macromol ; 193(Pt A): 585-591, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34699896

RESUMO

In recent years, Cas12a, a new member of the CRISPR family, has been found to have both DNase and RNase activities, have a simple structure, and a single promoter can simultaneously initiate multiple crRNAs, making the CRISPR/Cas12a editing system more advantageous in terms of structure and mechanism of action. Our team has successfully constructed Cas12a system that can be used in silkworm. Cas12a can be used to edit the multiple target sites. In production, a lot of factors can affect the production of silk industry. In order to make the silkworm resistant to the virus, using gene editing technology to knock out key genes for replication and proliferation in the Bombyx mori nuclearpolyhedrosisvirus (BmNPV) genome. Multiple sites on the BmNPV genome were selected as the target sites. We constructed the multi-sites expression vector of gie1-M (361 bp, 597 bp, 927 bp of ie-1) that edited multiple sites of BmNPV ie-1. The effects of multi-sites editing system on the proliferation and replication of the virus after the BmNPV genome was knocked out were examined. The results show that compared with CRISPR/FnCas12a single-site editing (gie1), multi-sites editing (gie1-M) can knock out the BmNPV genome more effectively and have a higher inhibitory effect on virus replication and proliferation. This system can provide a new direction for the breeding of silkworm resistant materials, and it can also lay a good technical platform for the identification and research of biological gene function.

9.
Spine J ; 21(11): 1881-1889, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34174436

RESUMO

BACKGROUND CONTEXT: Long-level spinal fusion for degenerative lumbar scoliosis (DLS)seeks to eliminate spinal motion in an attempt to alleviate pain, improve deformity, and reduce disability. However, this surgery considerably impairs the performance of activities of daily living (ADL) due to the resulting stiffness. The lumbar stiffness disability index (LSDI) is a validated measure of the effect of lumbar stiffness on functional activity, but this index might not be fully applicable to the elderly Chinese population given several specific lifestyle characteristics. PURPOSE: To evaluate lumbar stiffness in patients with DLS after long-level fusion by Chinese-LSDI (C-LSDI). STUDY DESIGN: A retrospective study. PATIENT SAMPLE: A total of 129 DLS patients who underwent long-level (≧4 levels) fusion surgery with at least one-year follow-up from June 2009 to September 2017 were retrospectively included. OUTCOME MEASURES: The C-LSDI was designed by modifying LSDI and Korean-LSDI (K-LSDI) based on elderly Chinese lifestyles and the internal consistency and retest repeatability of the patient-reported outcome questionnaire in the measurement of the impact of lumbar stiffness on functional abilities was assessed. METHODS: The radiographic parameters including Cobb angle, apical vertebral translation (AVT), coronal vertical axis (CVA), sagittal vertical axis (SVA), thoracic kyphosis (TK), thoracolumbar kyphosis (TLK), pelvic tilt (PT), pelvic incidence (PI), sacral slope (SS), lumbar lordosis (LL), and PI-LL, and clinical symptoms including visual analogue scale (VAS) for back and low extremity pain; Oswestry disability index (ODI), Japanese Orthopedic Association-29 (JOA-29), Scoliosis Research Society⁃22 (SRS-22), 36⁃Item short form survey (SF-36), physical component scores (PCS) and mental component scores (MCS) were measured preoperatively and at the last follow-up. RESULTS: Compared with LSDI and K-LSDI, the C-LSDI demonstrated higher internal consistency (Cronbach's alpha=0.902) and retest reliability (Internal consistency coefficients, ICC=0.904) in the elderly Chinese population. All patients showed increased lumbar stiffness and significant improvement in pain and deformity postoperatively. Regarding items, such as performing personal hygiene after toileting and getting out of a car, people reported more inconvenience with increasingly fixed levels. CONCLUSIONS: This study demonstrated that the C-LSDI questionnaire was a reliable and valid instrument for assessing functional limitations due to lumbar stiffness among elderly Chinese patients with DLS after long-level fusion. Although the effects of stiffness did trend toward greater impacts among patients who underwent longer fusions, most patients were satisfied with trade-offs of function and pain relief in exchange for perceived increases in lumbar stiffness.

10.
Int Urogynecol J ; 2021 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-34028577

RESUMO

INTRODUCTION AND HYPOTHESIS: The aim of the study was to develop artificial intelligence (AI) algorithms using 2D transperineal ultrasound (TPUS) static images to simplify the clinical process of diagnosing stress urinary incontinence (SUI) in practice. METHODS: The study involved 400 patients in total, including 265 SUI patients and 135 non-SUI patients who underwent a routine clinical evaluation process by urologists and TPUS. They were classified into different groups based on the International Consultation on Incontinence Questionnaire (ICIQ) to assess the impact of inconvenience on patients' lives. Four AI models were developed by 2D TPUS images: Model A (a single-mode model based on Valsalva maneuver images to classify G-0, G-1, and G-2); Model B (a dual-mode model based on Valsalva maneuver and resting state images to classify G-0, G-1, and G-2); Model C (a single-mode model based on Valsalva maneuver images to classify G-2 and G-01); Model D (a dual-mode model based on Valsalva maneuver and resting state images to classify G-2 and G-01). The performance of the four models was evaluated by confusion matrices and the area under the receiver-operating characteristic curve (AUC). RESULTS: The dual-mode model based on the Valsalva maneuver and resting-state images (Model D) had a higher accuracy of 86.3% and an AUC of 0.922, which was significantly higher than the AUCs of the other three models: 0.771, 0.862, and 0.827. CONCLUSIONS: The AI algorithm using 2D TPUS static images of the Valsalva maneuver and resting state may be a promising tool in the diagnosis of SUI patients in to relieve clinical processes in practice given its ease of use in clinical applications.

12.
Int Wound J ; 2021 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-33999495

RESUMO

Injuries and tumours of the cervical spine represent therapeutic challenges to the treating surgeon due to the complex anatomical relationships and biomechanical features. The anterior cervical midline (ACM) and anterior cervical retropharyngeal (ACR) approaches are effective and safe surgical approaches for certain cervical spine lesions, such as cervical spine neoplasms, atlantoaxial subluxation, and certain odontoid fractures. Posterior pharyngeal wall defects (PPWDs) is one of the most frequently encountered surgical morbidities after anterior cervical spine surgery (ACSS). However, limited information has been published concerning effective approaches for PPWD reconstruction after ACSS. The manuscript aimed to describe a novel application of the island sternocleidomastoid myocutaneous flap (ISMF) in the management of PPWDs after ACSS, including surgery with the ACM approach and ACR approach. From April 2015 to November 2019, the clinical data of three patients with PPWDs repaired using the ISMF in Peking university third hospital were retrospectively analysed. The observational indexes are as follows: postoperative survival of the flap, wound healing 2 weeks after surgery, eating and pronunciation function 2 months after surgery. The above indexes of these three cases recovered well. Three patients did not have any persistent PPWD after repair with the ISMF and did not require any further surgical procedures related to the cervical spine.

13.
Global Spine J ; : 21925682211017477, 2021 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-34018438

RESUMO

STUDY DESIGN: Retrospective study. OBJECTIVE: To investigate the incidence, risk factors, and outcomes of pedicle screw loosening in degenerative lumbar scoliosis (DLS) undergoing long-segment spinal fusion surgery. METHODS: One hundred and thirty DLS patients who underwent long-segment fusion surgery with at least a 12-month follow-up were studied. The incidence and risk factors of screw loosening were investigated. VAS, SRS-22, and ODI scores were obtained preoperatively and at follow-up. RESULTS: One hundred and sixty-eight of 1784 (9.4%) screws showed evidence of loosening in 71 (54.6%) patients. Three patients required revision surgery. Screw loosening rates according to vertebral insertion level were lowest instrumented vertebra (LIV): 45.4%; uppermost instrumented vertebra (UIV):17.7%; one vertebra above the LIV: 0.5%; 2 vertebrae above the LIV: 0.4%. Multiple logistic regression analysis of possible risk factors indicated that preoperative lateral subluxation ≥8 mm (odds ratio [OR]: 2.68, 95% confidence interval [CI]: 1.16-6.20), osteopenia (OR: 5.52, 95% CI: 1.64-18.56), osteoporosis (OR: 8.19, 95% CI: 2.40-27.97), fusion to sacrum (OR: 2.55, 95% CI: 1.12-5.83), postoperative TLK greater than 10° (OR: 2.63, 95% CI: 1.14-6.04) and SVA imbalance (OR: 3.44, 95% CI: 1.17-10.14) were statistically significant. No difference was noted in preoperative, follow-up, and change of VAS, ODI, and SRS-22 scores. CONCLUSIONS: Screw loosening in DLS underwent long-segment surgery is common and tends to occur in the LIV or UIV. Lateral subluxation ≥8 mm, osteopenia, osteoporosis, fusion to the sacrum, postoperative TLK greater than 10°, and SVA imbalance were the independent influencing factors. Screw loosening can be asymptomatic, while longer-term follow-up is required.

14.
Int Wound J ; 18(6): 796-804, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33733609

RESUMO

This study aimed to explore the treatment effect of Z-plasty on a non-healing wound. A total of 72 patients diagnosed with a chronic non-healing wound in Peking University Third Hospital from November 2009 to August 2019 were retrospectively analysed. Among them, 27 patients were treated with Z-plasty, and 45 patients were treated with the general method. Detailed patient information was retrieved from medical records, including age, gender, body mass index (BMI), alcohol, smoking, and comorbidities (diabetes mellitus, hypertension, heart disease). Surgical parameters included operation time and intraoperative blood loss. Wound swelling, epidermal blisters, wound edge colour, and skin temperature at 1 day after surgery were assessed to evaluate the blood supply of the wound. Surgical complications included infection, haematoma, dehiscence, and non-healing within 2 weeks postoperatively. Student t test (for continuous data) and Chi-square test (for categorical data) were conducted to determine the statistical difference. We found no significant differences in age, gender, BMI, alcohol, smoking, and comorbidities between the two groups. Z-plasty did not show any advantages in the surgical time, invasive blood loss, hospital days, and hospitalisation expenses. The incidence of abnormal wound edge colour with Z-plasty was significantly lower than that with the general treatment (P < .05), and the Z-plasty enables better healing of the patient's wound (P < .05). Z-plasty promoted better recovery of chronic non-healing wounds than direct suturing.


Assuntos
Transplante de Pele , Cicatrização , Vesícula , Hospitalização , Humanos , Estudos Retrospectivos
15.
Quant Imaging Med Surg ; 11(2): 706-713, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33532270

RESUMO

Background: There has been a long-standing controversy about diastasis recti diagnostic criteria and its relation to pelvic floor dysfunction (PFD). This study aimed to establish ultrasound diagnostic criteria for diastasis recti and investigate the correlation between it and PFD in early postpartum females. Methods: The inter-rectus distance (IRD) was measured at 3 locations in 116 healthy nulliparous females and 108 postpartum females. At the same time, they remained relaxed and then maintained a head-lift posture. The measurement for the 90th percentile was used to define the normal IRD in the nulliparous group. Afterward, the 108 postpartum females underwent an ultrasonographic examination of pelvic floor function. The correlations of these values with the IRD were then examined. Results: We established the following ultrasonographic diagnostic criteria for diastasis recti: an IRD of >2 mm at 3 cm below the umbilicus, >20 mm at the umbilicus, and >14 mm at 3 cm above the umbilicus. The IRD was positively correlated with body mass index (BMI) in the nulliparous group (r=0.286, P<0.01) and with age in the postpartum group (r=0.230, P<0.05). The IRD was not relative to either the PFD ultrasound results or the clinical symptoms. Conclusions: We established a set of ultrasonographic diagnostic criteria for diastasis recti at 3 locations along the umbilicus. There is no clear correlation between diastasis recti and PFD in early postpartum females.

16.
Eur Spine J ; 30(5): 1133-1143, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33210198

RESUMO

PURPOSE: To assess the lumbar muscle conditions on the incidence of proximal junctional kyphosis (PJK) after long-level correction and instrumentation surgery for degenerative lumbar scoliosis (DLS) patients with a minimum 2-year follow-up. METHODS: Eighty-four DLS patients undergoing long instrumented fusion surgery (≥ 5 vertebrae) were retrospectively studied. According to the occurrence of PJK at the final follow-up, patients were divided into the PJK group and the Non-PJK group. Patient characteristics, surgical variables and radiographic parameters were analyzed statistically. The lumbar muscularity (cross-sectional area of muscle-disc ratio × 100) and fatty degeneration (signal intensity of muscle-subcutaneous fat ratio × 100) were evaluated on magnetic resonance imaging . RESULTS: The prevalence of PJK was 20.24%. Gender, age at surgery, body mass index, uppermost instrumented vertebrae level, fusions extending to the sacrum, and levels fused were not significantly different between the groups. Lower bone mineral density, smaller functional cross-sectional area (FCSA) of paraspinal extensor muscles (PSE), higher lean muscle-fat index and total muscle-fat index of PSE, greater preoperative thoracolumbar kyphosis (TLK), smaller preoperative sacral slope (SS), larger preoperative sagittal vertical axis were identified in PJK group. Logistic regression analysis showed that osteoporosis, preoperative TLK > 15°, SS > 24°, FCSA of PSE > 138.75, and total muscle-fat index of PSE > 4.08 were independently associated with PJK. The final follow-up VAS score for back pain was higher, and SRS-22 subcategories of pain, function, self-image, and total score were significantly lower in the PJK group. CONCLUSION: Osteoporosis, lower lumbar muscularity and higher fatty degeneration, preoperative greater TLK and smaller SS were found to be strongly associated with the presence of PJK in DLS.


Assuntos
Cifose , Escoliose , Fusão Vertebral , Humanos , Vértebras Lombares , Complicações Pós-Operatórias , Estudos Retrospectivos , Fatores de Risco
17.
Sheng Wu Gong Cheng Xue Bao ; 37(12): 4342-4350, 2021 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-34984879

RESUMO

The CRISPR/Cas9 gene editing system has been widely used in basic research, gene therapy and genetic engineering due to its high efficiency, fast speed and convenience. Meanwhile, the discovery of novel CRISPR/Cas systems in the microbial community also accelerated the emergence of novel gene editing tools. CRISPR/Cpf1 is the second type (V type) CRISPR system that can edit mammalian genome. Compared with the CRISPR/Cas9, CRISPR/Cpf1 can use 5'T-PAM rich region to increase the genome coverage, and has many advantages, such as sticky end of cleavage site and less homologous recombination repair. Here we constructed three CRISPR/Cpf1 (AsCpf1, FnCpf1 and LbCpf1) expression vectors in silkworm cells. We selected a highly conserved BmHSP60 gene and an ATPase family BmATAD3A gene to design the target gRNA, and constructed gHSP60-266 and gATAD3A-346 knockout vectors. The efficiency for editing the target genes BmATAD3A and BmHSP60 by AsCpf1, FnCpf1 and LbCpf1 were analyzed by T7E1 analysis and T-clone sequencing. Moreover, the effects of target gene knockout by different gene editing systems on the protein translation of BmHSP60 and BmATAD3A were analyzed by Western blotting. We demonstrate the CRISPR/Cpf1 gene editing system developed in this study could effectively edit the silkworm genome, thus providing a novel method for silkworm gene function research, genetic engineering and genetic breeding.


Assuntos
Bombyx , Edição de Genes , Animais , Bombyx/genética , Bombyx/metabolismo , Sistemas CRISPR-Cas/genética , Endonucleases/genética , RNA Guia/genética
18.
Spine (Phila Pa 1976) ; 45(24): E1669-E1676, 2020 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-33231944

RESUMO

STUDY DESIGN: A retrospective study. OBJECTIVE: The aim of this study was to evaluate the factors affecting health-related quality of life (HRQOL) after surgery in patients with degenerative lumbar scoliosis (DLS) by minimum clinically important difference (MCID). SUMMARY OF BACKGROUND DATA: MCID has been introduced in the adult spinal deformity to quantify the absolute minimum change that can be considered a success. There are limited data available to identify factors affecting reaching MCID after DLS surgery. METHODS: This study reviewed a cohort of 123 DLS patients after correction surgery and with a minimum 2-year follow-up (FU). Inclusion criteria included age ≥40 and minimum five vertebrae fused and the availability of Scoliosis Research Society (SRS)-22 scores and radiographic data at baseline (BL) and FU. Using a multivariate analysis, two groups were compared to identify possible risk factors: those who reached MCID in the all four SRS domains (N = 65) at the last FU and those who missed MCID (N = 58). RESULTS: At baseline, patients differed significantly from matched normative data in all SRS-22 domains. The baseline HRQOL was comparable in reached MCID and missed MCID group patients (P > 0.05). The HRQOL scores at FU were significantly higher than those at baseline. Of 123 included patients, 77.2% (N = 95), 72.4% (N = 89), 76.4% (N = 94), and 89.4% (N = 110) reached MCID in SRS pain, activity, appearance, and mental domain, respectively. Pelvic incidence (PI) >55°, lumbar lordosis (LL) loss >4.65°, coronal imbalance at FU, sagittal vertical axis (SVA) at FU >80 mm, and presence of proximal junctional kyphosis (PJK) and distal junctional problem (DJP) had negative effects on the recovery process. CONCLUSION: Factors affecting reaching MCID after surgery for DLS were higher PI, LL loss, coronal imbalance, severe sagittal imbalance, and the occurrence of PJK and DJP. LEVEL OF EVIDENCE: 4.


Assuntos
Diferença Mínima Clinicamente Importante , Escoliose/cirurgia , Adulto , Idoso , Feminino , Humanos , Cifose/cirurgia , Lordose/cirurgia , Masculino , Pessoa de Meia-Idade , Qualidade de Vida , Estudos Retrospectivos , Fatores de Risco , Adulto Jovem
19.
Artigo em Inglês | MEDLINE | ID: mdl-32760714

RESUMO

CRISPR/Cas12a (Cpf1) is a single RNA-guided endonuclease that provides new opportunities for targeted genome engineering through the CRISPR/Cas9 system. Only AsCas12a has been developed for insect genome editing, and the novel Cas12a orthologs nucleases and editing efficiency require more study on insects. We compared three Cas12a orthologs nucleases, AsCas12a, FnCas12a, and LbCas12a, for their editing efficiencies and antiviral abilities. The three Cas12a efficiently edited the Bombyx mori nucleopolyhedrovirus (BmNPV) genome and inhibited BmNPV replication in BmN-SWU1 cells. The antiviral ability of the FnCas12a system was more efficient than that of the SpCas9 system after infection by BmNPV. We created FnCas12a × gIE1 and SpCas9 × sgIE1 transgenic hybrid lines and evaluated the gene-editing efficiency of different systems at the same target site. We improved the antiviral ability using the FnCas12a system in transgenic silkworm. This study demonstrated the use of the CRISPR/Cas12a system to achieve high editing efficiencies, and increase disease resistance in the silkworm.

20.
Nanomedicine ; 28: 102229, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32502696

RESUMO

As problems with the overuse of radical prostate cancer (PCa) treatment are increasingly exposed, focal therapy represents the direction of low- or intermediate-risk PCa management in the future. However, inaccurate diagnosis and low controllability of focal therapy hinder its clinical translation. In this study, we develop simple structural cyclic arginine-glycine-aspartic (cRGD) peptide-modified and indocyanine green (ICG)-loaded microbubbles (cRGD-ICG-MBs) for ultrasound-photoacoustic imaging and multi-synergistic photothermal therapy (PTT) to address the above problems. Precise PCa diagnosis is achieved by molecular ultrasound imaging. cRGD-targeting and low-frequency ultrasound with an amplitude of 500kPa convert MBs into nanoparticles for enhanced ICG delivery. Alow frequency2500 kPa amplitude ultrasound enables temporary vasculature destruction, which minimizes heat loss during PTT. Specifically, ICG in the tumor region is 14-fold higher than the control, resulting in satisfactory PTT. Our study highlights that this theranostic strategy possesses considerable clinical translational potential, especially in mini-invasive and individualized PCa therapy.


Assuntos
Verde de Indocianina/química , Terapia Fototérmica/métodos , Animais , Humanos , Masculino , Microbolhas , Técnicas Fotoacústicas/métodos , Neoplasias da Próstata/tratamento farmacológico , Nanomedicina Teranóstica/métodos
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