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Objective:To observe the effect of adipose tissue decellularized matrix hydrogel (DAT-gel) on the repair of sciatic nerve defect in rats.Methods:From April, 2019 to April, 2020, aseptic granular adipose tissue was collected from healthy adult women who underwent thigh or abdominal liposuction in the Department of Plastic Surgery, the First Medical Centre of the PLA General Hospital. Decellularisation and enzymatic digestion of adipose tissue were performed to prepare DAT-gel. Scanning electron microscope (SEM) was used to observe the ultrastructure of the hydrogel, and rheology was employed to test the gel dynamics and viscoelasticity of the hydrogel. A rat model of sciatic nerve defect was established and randomly divided into 3 groups: simple chitin catheter group (Chitin group), DAT-gel plus chitin catheter group (DAT-gel group) and autologous nerve reverse connection group (Autograft group) with 10 rats in each group. At the 12th week after surgery, the general view, function and morphology of the regenerated nerve were observed to evaluate the repairing status of the injured nerve. One-way analysis of variance (one-way ANOVA) was used for data analysis. If the difference between the groups was statistically significant, the Turkey method was further used for pairwise comparison. P<0.05 was considered as statistically significant. Results:The results of SEM showed that the DAT-gel had a three-dimensional structure in porous fibre network. The results of rheological test results showed that the complex viscosity of the hydrogel at 4 ℃ and 37 ℃ were 148.91 mPa·s and 801.29 mPa·s, respectively. DAT-gel underwent a sol-gel phase transition when the temperature had been increased. The results showed that DAT-gel had a good temperature-sensitive effect, and its critical point of sol-gel phase transition was similar to the internal temperature of rat. The results of animal experiments showed that the morphology and function of the regenerated nerve in the DAT-gel group were superior to Chitin group at 12 weeks after surgery, according to macroscopic view of the regenerated nerve, electrophysiology of the nerve, the morphology of the new axon and the target muscle, etc.. There was statistically significant between groups ( P<0.05). Conclusion:DAT-gel can significantly promote a repair of sciatic nerve defects in rats.
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We used CRISPR/Cas9 to delete plin1 of 3T3-L1 preadipocyte, to observe its effect on lipolysis in adipocytes and to explore regulatory pathways. We cultured 3T3-L1 preadipocytes, and the plin1 knockout vectors were transfected by electroporation. Puromycin culture was used to screen successfully transfected adipocytes, and survival rates were observed after transfection. The optimized "cocktail" method was used to differentiate 3T3-L1 preadipocytes. The glycerol and triglyceride contents were determined by enzymatic methods. The changes in lipid droplet form and size were observed by Oil red O staining. The protein expression of PLIN1, PPARγ, Fsp27, and lipases was measured by Western blotting. RT-PCR was used to measure the expression of PLIN1 and lipases mRNA. After the adipocytes in the control group were induced to differentiate, the quantity of tiny lipid droplets was decreased, and the quantity of unilocular lipid droplets was increased and arranged in a circle around the nucleus. Compared with the control group, the volume of unilocular lipid droplets decreased, and the quantity of tiny lipid droplets increased after induction of adipocytes in the knockout group. The expression of PLIN1 mRNA and protein in the adipocytes was significantly inhibited (P<0.05); glycerol levels increased significantly (0.098 4±0.007 6), TG levels decreased significantly (0.031 0±0.005 3); mRNA and protein expression of HSL and ATGL increased (P<0.05); PPARγ and Fsp27 expression unchanged in adipocytes. The above results indicate that the knockout of plin1 enhances the lipolysis of 3T3-L1 adipocytes by exposing lipids in lipid droplets and up-regulating lipases effects.
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Animals , Mice , 3T3-L1 Cells , Adipocytes , Metabolism , CRISPR-Cas Systems , Gene Knockout Techniques , Lipase , Metabolism , Lipolysis , Genetics , Perilipin-1 , Genetics , MetabolismABSTRACT
Objective To investigate the effect of Liensinine on lipopolysaccharide ( LPS)-induced acute lung injury (ALI) in mice. Methods BALB/c mice were randomly divided into six group: control group, LPS group, LPS+Liensinine (2 mg/kg, 4 mg/kg, 8 mg/kg) groups, and dexamethasone group. Acute lung injury in mice was induced by nasal instillation of LPS. After 12 h, the pathological changes of lung tissue were observed. The levels of TNF-α, IL-6 and IL-1β in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The number of neutrophils in BALF was detected using Wright-Giemsa staining. Total protein content was detected by BCA protein quantification assay. The pulmonary capillary permeability was examined with Evans blue. The MPO activity, MDA content, SOD activity, and GSH content in lung homogenate supernatant were detected by spectrophotometry. The content of ROS in lung tissue was detected by flow cytometry. Results The LPS group showed inflammatory cell infiltration, thickening of bronchial alveolar wall and pulmonary congestion in the lung tissue, while Liensinine improved the lung injury. In the LPS group, the contents of TNF- α, IL-6 and IL-1β in BALF were significantly increased, the number of neutrophils and the content of total protein were significantly increased, pulmonary capillary permeability, MPO activity and MDA content were increased, SOD activity and GSH content were decreased, the content of ROS was increased; while the Liensinine group reduced the contents of TNF-α, IL-6, IL-1β in BALF, reduced the number of neutrophils and total protein content, decreased the pulmonary capillary permeability, attenuated MPO activity and MDA contents and increased SOD activity and GSH content, and reduced ROS content in the LPS-challenged lung tissue. Conclusions Liensinine protects mice from LPS-induced acute lung injury by its anti-inflammatory and anti-oxidative activities.
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Objective To establish a stably overexpressing miR-31 transgenic mouse and detect the expression of miR-31 in the organs and tissues,and to provide qualified tool mice with overexpression of miR-31 in vivo. Methods The miR-31 overexpression vector was constructed by Gateway cloning technology. The vector was injected into fertilized ovum by DNA microinjection technology,then transferred to the pseudopregnant mice and waited for eutocia. Newborn mouse tail DNA was extracted and PCR and agarose gel electrophoresis were performed to identify the positive miR-31 transgenic mice. microRNA was extracted from the organs and tissues of miR-31 transgenic mice and the expression of miR-31 was de-tected by RT-PCR. The expression of Nestin and number of neural stem cells in the nervous system were compared in the positive and WT mice. Results The miR-31 transgenic mice were constructed successfully and bred more than 14 genera-tions in barrier environment. Expression of miR-31 was increased in major organs and tissues. The expression of Nestin and the number of neural stem cells in the positive mice were higher than those in the wild type mice. Conclusions MiR-31 overexpressing transgenic mice are constructed by Gateway cloning technology and the expression of miR-31 is stable in sub-sequent generations. The number of neural stem cells in the nervous system is higher than that in wild-type mice. The miR-31 overexpressing transgenic mice can be a good tool for experimental research of the function of overexpressed miR-31 in vivo and the treatment of nervous system diseases.
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Objective This study was designed to explore the therapeutic effect of psoralen on type Ⅱ collagen-induced rheumatoid arthritis in mice and its molecular mechanism.Methods DBA/1J mice were immunized with type II bovine collagen to induce rheumatoid arthritis.The model mice were randomly divided into Psoralen group(PSO),methotrexate group(MTX) and model group(Vehicle).Clinical signs of arthritis in the mice were monitored.The spleen index was assessed.Splenic Th1 and Th2 cells were counted by flow cytometry.ELISA was used to detect the levels of inflammation-associated factors TNF-α,IL-6 and IL-1β in the serum.Results Compared with the vehicle group,the ankle swelling and limitation of joint activity in the PSO group were significantly reduced,the spleen index and Th1 cell percentage were significantly decreased,and the Th2 cell percentage showed no significant change in the PSO group.Expression of TNF-α,IL-6 and IL-1β in serum was notably decreased in the PSO group.All the indexes showed no significant difference between the PSO and MTX groups.Conclusions Psoralen may attenuate the severity of type II collagen-induced rheumatoid arthritis in mice by regulating the balance of Th1/Th2 cells and inhibiting the expression of TNF-α,IL-6 and IL-1β.
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Objective@#To investigate the value of Multislice computed tomography volume rendering(VR) technique and 3D printing technique in auricular reconstruction.@*Methods@#Six patients were enrolled for auricular reconstruction with costal cartilage, including 5 congenital microtia patients and 1 traumatic auricular defect patient. We harvest the three-dimensional reconstructive data of the contralateral sixth, seventh, eighth and ninth costal cartilage with VR technique. Three-dimensional solid models were 3D printed with nylon material according to the data exported in STL format. Preoperative simulation was performed on the models, accordingly, we determined the strategies of costal cartilage harvest and framework fabrication, and operations were performed based on the pre-designed plan.@*Results@#In all 6 patients, the actual costal cartilage harvest and framework fabrication process was consistent with the preoperative design and simulation results, and more scientific than before. The shapes of reconstructed ears were vivid and natural. No complications such as infection, absorption, distortion and chest deformity happened.@*Conclusions@#Through costal cartilage VR and 3D printing technique, we could make more reasonable preoperative design and simulation. The results can be improved with reduced injury, while avoiding the risks of thoracic deformity.
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Chinese hamster is an important laboratory animal in medical and biological researches,but the molecular genetic markers research was rarely reported.In our study the base composition,gene structure,genetic evolution and other characteristics of mitochondrial genome of Chinese hamster were analyzed using the methods of bioinformatics and comparative genomics,genetic quality detection system of Chinese hamster were also established.These results would supply genome data for animal models of human diseases,and lay the foundation for scientific evaluation and reasonable utilization.
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Objective To study the expression and significance of EGFR, PDCD4, TGF-β1, Smad3, Smad7 in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia and oral squamous cell carcinomas(OSCC).Methods Serum levels of EGFR, PDCD4 were measeured with ELISA in Chinese hamster during the oral mucosa carcinogenesis;The expressions of TGF-β1, Smad3, Smad7 in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia and OSCC tissues were examined by immunohistochemistry.Results In the process of oral carcinogenesis,the expression level of EGFR increased significantly,while the expression of PDCD4 was decreased, the negatively correlation was evident between expression of these two proteins in ED and SCC.The expression of TGF-β1, Smad7 was higher in OSCC than in oral normal mucosa, oral simple hyperplasia, oral epithelial dysplasia(P < 0.05), while the expression of Smad3 was decreased.Further analysis showed the expression of TGF-β1 was correlated with the expression of Smad7.Conclusions The expressions of EGFR, PDCD4, TGF-β1, Smad3, Smad7 are closely related with the biological behaviors of oral squamous cell cancinoma.This experiment provides a theoretical basis for the study of oral squamous cell carcinomas mechanism research.So it has important significance for the development and prognosis of oral squamous carcinomaare.
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Objective To investigate the effect of uterine cavity injection of absolute ethanol on estrous cycle in mice.Methods Twenty mice with regular estrous cycle were selected by vaginal exfoliated cells staining, and then were injected absolute ethanol into their uterine cavity.The estrous cycle was observed and recorded every day.Ten mice were selected randomly to observe the changes of second estrous cycle every two hours.Results After the injection of absolute ethanol into the uterine cavity of the experimental mice, the estrous cycle was greatly prolonged, and the various degree of disorder was observed in estrus and pre-estrus.Conclusions The disorder was observed in estrous cycle of mice after the injection of absolute ethanol, which maybe provides some references for clinical use of absolute ethanol.
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Objective To report our first clinical experience with a novel modified culotte technique for the treatment of true coronary bifurcation lesions. Methods The novel modified culotte technique (the mono-ring culotte) stenting was done in which the side branch (SB) stent was deployed firstly followed by ex vivo wiring of a most proximal cell of SB stent with the hard end of main branch (MB) wire. Secondly, the MB stent was deployed through the most proximal cell of SB stent. The procedure was ended with kissing balloon dilation. From June 2014 to March 2015, 15 patients with true coronary bifurcation lesion were treated with mono-ring culotte stenting in our center. Results The procedures were successful in all cases without procedural complication and in-hospital major adverse cardiovascular events. The procedural time was (34. 3 ± 9. 6) min, fluoroscopic time was (18. 1 ± 3. 8) min, and contrast volume was (112. 0 ± 24. 5) ml, respectively. Post-procedurally, the residual stenosis of the main and the side branch were (10. 0 ± 2. 5)% and (10. 2 ± 5. 3)% , respectively. Conclusions The mono-ring culotte stenting is safe and feasible for treatment of true coronary bifurcation lesions, and may be superior to the conventional culotte stenting.
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<p><b>OBJECTIVE</b>To investigate the role of connexin 43-formed hemichannels in cell volume regulation induced by simulated ischemia/reperfusion (SI/R).</p><p><b>METHODS</b>Mouse cardiomyocytes isolated on a Langendorff apparatus with enzyme solution were aliquoted into control, SI/R and SI/R +octanol groups. Calcein-AM was used to stain the cells and the cell volume was measured with confocal microscope by stack scanning. Trypan blue was used to measure the cell viability after the treatments.</p><p><b>RESULTS</b>Calcein-AM staining and cofocal microscopy yielded stable and reproducible results for cell volume measurement. Mouse cardiomyocytes subjected to simulated SI/R showed obvious cell swelling as compared with the control cells [(126∓6)% vs 100%, P<0.05], and octanol preconditioning significantly attenuated the cell swelling [(113∓6)%, P<0.05]. SI/R caused a significant reduction of the cell viability compared to the control cells [(19∓2)% vs (45∓3)%, P<0.01], and octanol preconditioning obviously reduced the viability of the cells with SI/R challenge [(31∓2)%, P<0.01].</p><p><b>CONCLUSION</b>Connexin 43-formed hemichannels are involved in the regulation of cardiomyocyte volumes induced by SI/R challenge, and octanol can alleviate the cell swelling to enhance the viability of the cardiomyocytes following SI/R.</p>
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Animals , Mice , Cell Line , Cell Size , Cell Survival , Connexin 43 , Metabolism , Ischemic Preconditioning, Myocardial , Methods , Mice, Inbred C57BL , Myocardial Reperfusion Injury , Metabolism , Pathology , Myocytes, Cardiac , Pathology , Octanols , PharmacologyABSTRACT
To investigate the safety, feasibility and effectiveness of transrectal high-intensity focused ultrasound (HIFU) in the ablation of canine prostate, 20 dogs were divided randomly into 5 groups. Sixteen canine prostates were treated with the third-generation transrectal HIFU device (Sonablate-500). Transrectal ultrasound images of the prostate and prostatic urethra were observed preoperatively and postoperatively. Serial study was performed 30 min, 30 days, 60 days and 180 days after the therapy. The rectum, periprostatic tissues, and prostate were excised en bloc and the tissues were fixed for gross and histological analysis. Our results showed that the average maximal diameter of prostatic urethra was 0.59+/-0.11 cm before the operation and 2.57+/-0.98 cm 60 days after the operation. The volume of prostate was 6.5+/-3.12 cm(3) before the treatment while the volume was 4.13+/-0.23 cm(3) 60 days after the treatment and the differences were statistically significant (P<0.05). Histologically, there was a clear demarcation between the necrotic area of the treated tissues and the unaffected surrounding tissues. All the necrotic tissues in the targeted zone broke off and the prostatic urethra became cavitary 60 days later. The more frequent complications were urinary retention and frequency and hematuria. No rectal injury occurred during the treatment. It is concluded that the third-generation transrectal HIFU is capable of destroying prostatic tissue, substantially increasing the width of the prostatic urethra without causing injury to the adjacent tissues. The risk of postoperative complications associated with HIFU was low. HIFU may become a safe, effective and minimally invasive alternative for the treatment of prostatic diseases.
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To investigate the safety, feasibility and effectiveness of transrectal high-intensity focused ultrasound (HIFU) in the ablation of canine prostate, 20 dogs were divided randomly into 5 groups.Sixteen canine prostates were treated with the third-generation transrectal HIFU device (Sonablate-500 TM). Transrectal ultrasound images of the prostate and prostatic urethra were observed preoperatively and postoperatively. Serial study was performed 30 min, 30 days, 60 days and 180 days after the therapy. The rectum, periprostatic tissues, and prostate were excised en bloc and the tissues were fixed for gross and histological analysis. Our results showed that the average maximal diameter of prostatic urethra was 0.59±0.11 cm before the operation and 2.57±0.98 cm 60 days after the operation. The volume of prostate was 6.5±3.12 cm3 before the treatment while the volume was 4.13±0.23 cm3 60 days after the treatment and the differences were statistically significant (P＜0.05).Histologically, there was a clear demarcation between the necrotic area of the treated tissues and the unaffected surrounding tissues. All the necrotic tissues in the targeted zone broke off and the prostatic urethra became cavitary 60 days later. The more frequent complications were urinary retention and frequency and hematuria. No rectal injury occurred during the treatment. It is concluded that the third-generation transrectal HIFU is capable of destroying prostatic tissue, substantially increasing the width of the prostatic urethra without causing injury to the adjacent tissues. The risk of postoperative complications associated with HIFU was low. HIFU may become a safe, effective and minimally invasive alternative for the treatment of prostatic diseases..
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To evaluate the safety and efficacy of transrectal high-intensity focused ultrasound (HIFU) in the treatment of benign prostatic hyperplasia (BPH), serial studies were conducted in 150 BPH pa- tients before and 30 min, 1, 2, 6 and 12 month(s) after Sonablate-500TM HIFU treatment. A sili- con-coated indwelling 16F latex catheter was placed during the determination of the therapy zone. Preoperative and postoperative evaluations were made by using the international prostate symptom score (IPSS), quality of life (QOL), uroflowmetric findings and transrectal ultrasound, and incidence of complications. The cystourethrography was done in 23 patients within 1 year postoperatively. The results showed that after HIFU treatment, IPSS and QOL scores were significantly decreased at 1, 2, 6 and 12 month(s) (P<0.01). Maximum urine flow rate (6.0 to 17.2 mL/s, P<0.01), PVR (75.0 to 30.3,P<0.01) and prostatic volume (65.0 to 38.1 mL, P<0.05) were significantly improved 12 months after the operation. Recurrent urinary retention (n=2) and urethrorectal fistula (n=1) occurred at the 15th postoperative day. The duration of the HIFU prostate ablation was 25-90 rain. The mean time for an indwelling catheter was 3-19 days. These data demonstrate that treatment of BPH with Sonab- late-500TM HIFU is safe and effective.