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1.
Journal of Medical Biomechanics ; (6): E052-E058, 2023.
Article in Chinese | WPRIM | ID: wpr-987913

ABSTRACT

Objective To verify the biomechanical stability of oblique lateral interbody fusion ( OLIF) combinedwith different fixation methods for treating degenerative lumbar scoliosis (DLS) by three-dimensional (3D) finite element analysis. Methods The L1-S1 3D finite element DLS model ( Model 1) was established, and then the OLIF (L2-5) at 3 contiguous levels of fusion and its combination with different internal fixation methods were simulated, namely, stand-alone OLIF model ( Model 2), vertebral screw fixation model ( Model 3), unilateral pedicle screw fixation model (Model 4) and bilateral pedicle screw fixation model (Model 5) were established,respectively. Under upright, flexion, extension, lateral bending and axial rotation states, range of motion (ROM) of fusion segments, as well as cage stress, internal fixation stress, and stress distribution were recorded and analyzed. Results Under six motion states, the overall ROM of fusion segments in Models 2-5 was smaller than that of Model 1. Compared with Model 1, the overall ROM reduction of Model 3 and Model 4 was larger than that of Model 2 and smaller than that of Model 5. Under flexion and extension, the overall ROM reduction of Model 4 and Model 5 was basically equal. Under left and right lateral bending, the overall ROM reduction of Model 3 and Model 5 was basically equal. Under all motion states, the peak stress of Model 3 and Model 4 fusion cage was larger than that of Model 5 and smaller than that of Model 2. The peak stresses of L2-3, L3-4 and L4-5 fusion cages in Model 3 increased by 5. 52% , 10. 96% and 7. 99% respectively compared with Model 5 under left lateral bending, and the peak stresses of L2-3, L3-4 and L4-5 fusion cages in Model 4 increased by 8. 70% , 7. 00% and 6. 99% respectively under flexion. Under all motion states, the peak stress of screw rod in Model 5 was smaller than that of Model 3 and Model 4, and the peak stresses of screw rod in Models 3-5 were the smallest in upright state. Conclusions The OLIF with unilateral pedicle screw fixation or vertebral screw fixation can provide favorable biomechanical stability of the fusion segment. The results provide some references for clinical application of OLIF technology in the treatment of DLS.

2.
Chinese Medical Journal ; (24): 1513-1522, 2023.
Article in English | WPRIM | ID: wpr-980939

ABSTRACT

Asthma is characterized by chronic airway inflammation and airway hyper-responsiveness. However, the differences in pathophysiology and phenotypic symptomology make a diagnosis of "asthma" too broad hindering individualized treatment. Four asthmatic inflammatory phenotypes have been identified based on inflammatory cell profiles in sputum: eosinophilic, neutrophilic, paucigranulocytic, and mixed-granulocytic. Paucigranulocytic asthma may be one of the most common phenotypes in stable asthmatic patients, yet it remains much less studied than the other inflammatory phenotypes. Understanding of paucigranulocytic asthma in terms of phenotypic discrimination, distribution, stability, surrogate biomarkers, underlying pathophysiology, clinical characteristics, and current therapies is fragmented, which impedes clinical management of patients. This review brings together existing knowledge and ongoing research about asthma phenotypes, with a focus on paucigranulocytic asthma, in order to present a comprehensive picture that may clarify specific inflammatory phenotypes and thus improve clinical diagnoses and disease management.


Subject(s)
Humans , Asthma/drug therapy , Inflammation/diagnosis , Respiratory System , Phenotype , Biomarkers , Sputum , Eosinophils , Neutrophils
3.
Chinese Journal of Tissue Engineering Research ; (53): 4547-4554, 2020.
Article in Chinese | WPRIM | ID: wpr-847309

ABSTRACT

BACKGROUND: Diffusion tensor imaging, as a relatively new method based on MRI, has become an important means of examination and diagnosis in the field of neuroimaging. OBJECTIVE: To investigate the role of using diffusion tensor tensor imaging data to predict 3D-bioprinted collagen/silk fibroin scaffolds in the locomotor function recovery after spinal cord injury. METHODS: Ordinary and 3D-bioprinted collagen/silk fibroin scaffold were prepared. Forty adult female SD rats provided by the Laboratory Animal Center of the Academy of Military Medical Sciences of the People’s Liberation Army were randomly divided into four groups with 10 rats in each group. In the sham operation group, only T10 vertebral plate was removed. In the model group, spinal cord injury was induced by total transection of spinal cord at T10 segment. In the ordinary collagen scaffold and 3D-printed scaffold groups, after induction of T10 spinal cord injury, ordinary collagen scaffold and 3D-printed scaffold were implanted, respectively. At 1, 2, 3, 4, 6 and 8 weeks after surgery, Basso, Beattie and Bresnahan (BBB) locomotor function scoring and oblique plate test of the hind limbs were carried out. At 8 weeks after surgery, electrophysiological test of the hind limbs was performed to evaluate locomotor function. At 8 weeks after surgery, diffusion tensor imaging of the lumbar spine was performed and the correlation between diffusion tensor imaging parameter and rat locomotor function was analyzed. Animal experiments were approved by the Animal Ethics Committee of Characteristic Medical Center of the Chinese people’s Armed Police Force (approval No. 27653/58). RESULTS AND CONCLUSION: (1) From 3 weeks after surgery, BBB score in the 3D-printed group was significantly higher than that in the model and ordinary collagen scaffold groups (P < 0.05 or P < 0.01). From 2 weeks after surgery, the slope angle in the 3D-printed scaffold group was significantly higher than that in the model and ordinary scaffold groups (P < 0.05 or P < 0.01). (2) The amplitude of motor evoked potential in the 3D-printed scaffold group was significantly greater than that in the model and ordinary collagen scaffold groups (P < 0.05 or P < 0.01). The latency of motor evoked potential in the 3D-printed scaffold group was significantly shorter than that in the model and ordinary collagen scaffold groups (P < 0.05 or P < 0.01). (3) Diffusion tensor imaging showed that the nerve fiber trajectories in the three groups were irregular and lacked the continuity of nerve fibers, but the number of regenerated nerve fiber bundles in the 3D-printed collagen scaffold group was greater than that in the model and ordinary collagen scaffold groups (P < 0.01). The fractional anisotropy at 9, 7.5, 4.5, -3, -6, -7.5, -9 mm from the center of spinal cord injury in 3D-printed collagen scaffold group was significantly higher than that in model and ordinary collagen scaffold groups (P < 0.05 or P < 0.01). (4) The BBB score, slope angle, amplitude of motor evoked potential, latency of motor evoked potential were positively correlated with the fractional anisotropy value of diffusion tensor imaging from head to tail of rats. (5) These results suggest that diffusion tensor imaging can be used as an effective predictor to evaluate the recovery of neurological function after spinal cord injury in experimental animals and clinical cases.

4.
Journal of Leukemia & Lymphoma ; (12): 570-573, 2020.
Article in Chinese | WPRIM | ID: wpr-862878

ABSTRACT

Diffuse large B-cell lymphoma (DLBCL) is the most common invasive non-Hodgkin lymphoma, which responds well to R-CHOP regimen immunochemotherapy, but 30%-40% of the patients eventually develop relapsed and refractory DLBCL. Therefore, the discovery of new prognostic markers is essential to improve the diagnosis and treatment of DLBCL. Immune escape is an important mechanism for the occurrence and development of DLBCL. Studies have shown that TIM-3, BTLA and LAG-3 are highly expressed in DLBCL, which could inhibit the effective function of immune cells in tumor microenvironment, and promote the immune escape of lymphoma cells. Thus, they promote the occurrence and development of DLBCL and affect the effect of conventional chemotherapy. It is important to explore the effects of the above three inhibitory molecules on DLBCL, and they are expected to become new targets for the treatment of DLBCL.

5.
Chinese Journal of Behavioral Medicine and Brain Science ; (12): 1057-1063, 2019.
Article in Chinese | WPRIM | ID: wpr-800493

ABSTRACT

Objective@#To explore the inhibitory effect of exosomes secreted by human umbilical cord mesenchymal stem cells(HUCMSC) on apoptosis of human umbilical vein endothelial cells(HUVEC) after model group(oxygen-glucose deprivation reoxygenation), and to clarify its possible mechanism.@*Methods@#Human umbilical cord mesenchymal stem cells were cultured. The collected cell supernatant was stored in a centrifugal tube. The exosomes secreted by human umbilical cord mesenchymal stem cells were extracted by ultracentrifugation and identified. Human umbilical vein endothelial cells were randomly divided into control group, model group and different concentrations of HUCMSC-EXO(20 μg/ml, 40 μg/ml, 60 μg/ml) treatment groups(adding HUCMSC-EXO into the model group) . The morphological changes of HUVEC cells in each group were observed by inverted phase contrast microscope, and the proliferation inhibition rate of HUVEC in each group was measured by CCK-8 reagent. Western blot was used to detect the expression of apoptosis-related proteins Caspase-3, Bax, Bcl-2 and hypoxia-associated protein hypoxia inducible factor 1α(HIF-1α). Inhibitor(HIF-1α inhibitor) + model group and HUCMSC-EXO + inhibitor + model group were added on the basis of the above experiments. Western blot analysis was performed to observe the effects of HUCMSC-EXO, inhibitor and both of them on HIF-1α and Bax expressions in HUVEC.@*Results@#HUCMSC-EXO was successfully extracted and identified. Compared with the control group, the volume of HUVEC in the model group and the HUCMSC-EXO group with different concentrations decreased, became round, connected and evacuated, and the growth state was poor under the inverted phase contrast microscope.CCK-8 detection showed that the cell viability in the HUCMSC-EXO group was significantly higher than that in the model group, the difference was statistically significant (t=9.23, P<0.05). Western blot analysis showed that compared with the control group, the expression levels of Caspase-3 ((0.296±0.038), (0.879±0.088); t=14.92, P<0.05), Bax((0.234±0.034), (0.762±0.084); t=14.36, P<0.05) of HUVEC in the model group were up-regulated, and the expression level of Bcl-2 was down-regulated ((0.863±0.103), (0.387±0.059); t=9.85, P<0.05), with statistically significant differences. Compared with the model group, the expression levels of Caspase-3( (0.586±0.075); t=6.24, P<0.05), Bax((0.311±0.055); t=11.01, P<0.05) and Bcl-2((0.665±0.071); t=7.45, P<0.05) of HUVEC in the HUCMSC-EXO treatment group were down-regulated and the differences were statistically significant. Inhibitor intervention experiments showed that there were no significant differences between the inhibitor+ model group and HUCMSC-EXO+ inhibitor+ model group in the expression of HIF-1α protein ((0.348±0.055), (0.388±0.077); t=1.04, P>0.05)and Bax protein ((0.363±0.069), (0.370±0.064); t=0.18, P>0.05). But both of them were down-regulated compared with the model group (HIF-1α protein (0.919±0.064), Bax protein (0.902±0.071)), the differences were significant( t=13.56, t=13.03, both P<0.05).@*Conclusion@#HUCMSC-EXO has a protective effect on OGD/R model of HUVEC, and its mechanism may be related to the down-regulation of HIF-1α expression.

6.
Chinese Journal of Ultrasonography ; (12): 691-695, 2019.
Article in Chinese | WPRIM | ID: wpr-754860

ABSTRACT

To assess the value of endorectal ultrasonography ( ERUS ) in predicting the pathological response to neoadjuvant chemoradiotherapy( NCRT ) for locally advanced rectal cancer( LARC) . Methods Ninety‐nine patients with LARC received NCRT and total mesorectal excision in our hospital were retrospectively analyzed . T he maximum length and thickness of rectal tumor were measured by ERUS both before NCRT ( ERUS1 ) and after NCRT following sugery ( ERUS2 ) ,and the length and thickness reduction rate were calculated . T he patients were classified into good responder group ( n = 47 ) and poor responder group( n = 52 ) ,or pathological complete response ( pCR) group ( n = 25 ) and non‐pCR group ( n=74) according to pathological tumor regression grade ( T RG ) . T he differences of various parameters were compared between groups . T he correlations between these parameters and T RG grading were analyzed by Spearman correlation analysis . T he ROC curve was used to evaluate the diagnostic efficacy of the parameter . Results T he length and thickness of ERUS2 were significantly shorter than that of ERUS1( all P <0 .05) . T he length and thickness of ERUS2 in good responder group were shorter than those in poor responder group ,while the length and thickness reduction rate were higher than those in poor responder group with significant difference ( all P < 0 .05 ) . T he length and thickness of ERUS2 in pCR group were shorter than those in non‐pCR group ,w hile the length and thickness reduction rate were higher than those in non‐pCR group with significant difference ( all P < 0 .05 ) . T he length and thickness of ERUS2 were positively correlated with T RG grading ( r = 0 .577 ,0 .605 ; all P < 0 .01 ) and the length and thickness reduction rate were negatively correlated with T RG grading ( r = -0 .681 ,-0 .598 ; all P <0 .01 ) . ROC curve showed the cut‐off value of the length and thickness reduction rate to predict good responder were 41 .34% ,46 .46% , with corresponding AUC areas of 0 .843 ,0 .796 , sensitivity of 74 .5% ,70 .2% , and specificity of 76 .9% ,80 .8% ,respectively . ROC curve showed the cut‐off value of the length and thickness reduction rate to predict pCR were 57 .36% ,58 .52% ,with corresponding AUC areas of 0 .851 and 0 .895 , sensitivity of 68 .0% ,76 .0% ,and specificity of 94 .6% ,93 .2% ,respectively . Conclusions T he changes of length and thickness of tumor after NCRT are well correlated with treatment response . T he length and thickness reduction rate measured on ERUS present high accuracy in prediction of good response and pCR in LARC patients .

7.
Chinese Journal of Cerebrovascular Diseases ; (12): 63-67, 2018.
Article in Chinese | WPRIM | ID: wpr-702989

ABSTRACT

Objective To compare the incidence of chronic subdural hematoma(CSDH) and its risk factors after surgical clipping unruptured intracranial aneurysms(UIA) and ruptured intracranial aneurysms (RIA). Methods From January 2006 to December 2015,410 consecutive patients with UIA and 464 patients with RIA treated with aneurysm clipping at the Department of Neurosurgery,Hanzhong Central Hospital were enrolled retrospectively. According to whether having postoperative CSDH or not(evaluating subdural effusion and its degree with head CT scan),the difference of the incidence of CSDH after aneurysm clipping in patients with UIA and RIA were compared,and logistic regression analysis was used to evaluate the risk factors for the occurrence of postoperative CSDH. Results (1) The incidence of CSDH after UIA clipping was higher than that of RIA(11.0% [45/410] vs. 3.0% [14/464]). There was significant difference (P<0.01). The rate of surgical treatment for CSDH because of the symptoms of nervous system injury in patients with UIA was higher than that in patients with RIA(35.6% [16/45] vs. 28.6% [4/14], P<0.05). (2) Multivariate logistic regression analysis showed that unruptured aneurysms (OR,2.59, 95% CI 2.19-3.06,P<0.01),subdural effusion ≥5 mm (OR,1.98,95% CI 1.94-2.03,P<0.01), and CT value≥40 HU (OR,2.87,95% CI 2.65-3.01,P<0.01) were the independent risk factors for postoperative CSDH in patients with intracranial aneurysm. Conclusions The incidence of CSDH was significantly higher than that of RIA after UIA clipping. UIA,subdural effusion ≥5 mm,and CT value ≥40 HU were the independent risk factors for CSDH of intracranial aneurysms.

8.
Chinese Journal of Geriatric Heart Brain and Vessel Diseases ; (12): 282-285, 2018.
Article in Chinese | WPRIM | ID: wpr-709113

ABSTRACT

Objective To study the value of CAS and brain atrophy index (BAD in predicting CSDH in elderly patients.Methods Two hundred and sixty-eight elderly head trauma patients were divided into CSDH group (n=40) and CSDH-free group (n=228).Their CSF volume,intracranial volume,brain volume,ventricular volume,subarachnoid space volume were measured by MR scanning,their BAI,Evans index,FHI,CAS score and clinical data such as gender and age were recorded,their clinical data and laboratory parameters were compared.Relationship between different factors in elderly CSDH patients was analyzed.The risk factors for CSDH in head trauma patients were analyzed by univariate and multivariate logistic regression analysis.Results The BAI and CAS score of moderate and severe brain atropy were significantly higher in CSDH group than in CSDH-free group (P<0.01).Correlation analysis showed that BAI was negatively related with brain volume and positively related with ventricular volume and subarachnoid space volume,ventriclular volume was positively related with subarachnoid space volume and Evans index,subarachnoid space volume was positively related with Evans index (r=0.35,P=0.02),Evans index was positively related with FHI (r=0.23,P =0.03).Univariate and multivariate logistic regression analysis showed that BAI≥7.15% on admission and CAS score of moderate and severe brain atrophy increased the risk of CSDH in elderly head trauma patients (P<0.05).Concluson CAS score and BAI can predict the occurrence of CSDH in elderly head trauma patients.

9.
Chinese Journal of Medical Ultrasound (Electronic Edition) ; (12): 411-416, 2017.
Article in Chinese | WPRIM | ID: wpr-711998

ABSTRACT

Objective To investigate the value of endorectal ultrasonography (ERUS) inpreoperative assessment of rectal cancer post neoadjuvant chemoradiation therapy.Methods From Jan.2016 to Dec.2016,90 rectal cancer patients who underwent preoperative neoadjuvant chemoradiation therapy and total mesorectal excision surgery in the Sixth Affiliated Hospital of Sun Yat-Sen University were retrospectively analyzed,and all patients underwent ERUS examination post neoadjuvant chemoradiation therapy.Of these,64 patients were evaluated by ERUS pre and post neoadjuvant chemoradiation therapy and 26 patients were evaluated only post neoadjuvant chemoradiation therapy.Wilcoxon rank sum test for paired sample was performed to compare the distance from inferior margin of tumor to anal margin,the length and thickness of the tumor pre and post neoadjuvant chemoradiation therapy respectively in rectal cancer.Taken pathologic findings as golden standard,the accuracy of T staging assessed by ERUS post neoadjuvant chemoradiation therapy was evaluated.Results Compared with pre neoadjuvant chemoradiation therapy,the distance from inferior margin of tumor to anal margin significantly increased after neoadjuvant chemoradiation therapy [(58.63±21.71) mm vs (51.68± 19.81) mm],and the length [(26.10± 10.07) mm vs (40.82±9.18) mm] and thickness [(9.73±2.50) mm vs (14.92±5.30) mm] of tumor also evidently decreased post neoadjuvant chemoradiation therapy,respectively (Z were 4.996,6.153 and 6.076,all P < 0.01).The final pathological T stage was pathologic complete response (pCR) or pT0 in 15 patients,pT1 in 3 patients,pT2 in 30 patients and pT3 in 42 patients.The diagnostic accuracy of T staging of rectal cancer post neoadjuvant chemoradiation therapy for ERUS was uT0 82.2% (74/90),uT1 96.7% (87/90),uT2 66.7% (60/90),uT3 67.8% (61/90) and uT4 96.7% (87/90),and the overall accuracy was 82.2% (74/90).Conclusion ERUS could effectively record the morphological changes of rectal cancer pre and post neoadjuvant chemoradiation therapy,which may contribute to the re-evaluation of the distance from inferior tumor margin to anal margin and the range and depth of tumor involvement pre surgical resection.

10.
Chinese Journal of Medical Imaging Technology ; (12): 1357-1361, 2017.
Article in Chinese | WPRIM | ID: wpr-607790

ABSTRACT

Objective To evaluate the value of endorectal ultrasonography (ERUS) in assessment of mesorectal fascia (MRF) invasion in rectal cancer.Methods Data of 44 patients who accepted preoperative ERUS and total mesorectal excision surgery within a week were retrospective analyzed.There were 18 patients who accepted preoperative neoadjuvant chemotherapy and 26 patients didn't acceped.Taking the pathological diagnosis of circumferential resection margin (CRM) as the gold standard,the diagnostic efficiency of ERUS for the MRF invasion in rectal cancer was evaluated.Results The final pathological T staging was T1 in 2 cases,T2 in 17 cases and T3 in 25 cases.There were 2 cases of CRM positive results,and 42 cases of CRM negative results.With regard to the location of tumor,there were 16 cases located in low,and 28 cases in mid rectum.There were 26 cases located in anterior or antero-lateral wall of rectum,13 cases in posterior or postero-lateral wall,and 5 cases with a circle of rectum.The diagnostic accuracy were 83.33 % (15/18) and 92.31% (24/26) for cases of accepting and not accepting the preoperative neoadjuvant chemotherapy;80.77% (21/26) for cases located in anterior or antero-lateral wall,and 100% (13/13) for cases located in posterior or postero-lateral wall;75.00% (12/16)and 96.43 % (27/28) for low position and mid position tumors.The total diagnostic accuracy was 88.64% (39/44).Conclusion ERUS can be an effective method in preoperative assessment of the MRF invasion in rectal cancer.

11.
Chinese Journal of Medical Ultrasound (Electronic Edition) ; (12): 467-472, 2015.
Article in Chinese | WPRIM | ID: wpr-637461

ABSTRACT

ObjectiveTo assess the diagnostic value of ultrasonography in the diagnosis and pathological classifi cation of appendicitis with pathological results.MethodsThe study included 111 cases who confi rmed by surgery and pathology in the Six Affi liated Hospital of Sun Yat-sen University from June2012 to December 2013. The image characteristics of ultrasonic images performed before surgery were analyzed retrospectively. The cases were divided into five groups based on the pathology: 6 acute simple appendicitis, 45 acute suppurative appendicitis, 15 acute gangrenous appendicitis, 33 chronic appendicitis, and 12 acute attack of chronic appendicitis. The length, width and wall thickness of the appendicitis measured by preoperative ultrasound between various pathological type were compared with Kruskal-Wallis statistical methord, and also used the same method to compared the one which had statistically signifi cance between the fi ve groups. As in the above case, Fisher Exact Test was used to compare the characters′ of ultrasonography included gradations of the appendix, the tube cavity expansion, stercorolith, the appendiceal abscess, mergering lymph node enlargement and the appendix around effusion, if there was statistically signifi cance, the same methord was used to the further comparison between thefi ve groups.ResultsTotally 90 of 111 cases of were reminded appendicitis by preoperative ultrasound, the diagnostic accuracy rate was 81.08%, including acute suppurative appendicitis 91.11% (41/45), acute gangrenous appendicitis 93.33% (14/15), acute simple appendicitis 83.33% (5/6), chronic appendicitis 60.61% (20/33), chronic appendicitis onset acute 83.33% (10/12). There were no statistical differences of preoperative ultrasound measured between various pathological type of appendicitis in length, while the appendix width measured by preoperative ultrasound of acute gangrenous appendicitis was wider than the acute simple group and chronic group, and there were statistically signifi cance between them (P<0.05), in the aspect of wall thickness, the acute suppurative appendicitis and the acute gangrenous appendicitis groups were thicker than the chronic group, and these differences had statistically signifi cance (P<0.05). The stercorolith, lymph node enlargement and the appendiceal abscess in different types of appendicitis had no statistical difference. There were statistically differences (P<0.05) between the acute suppurative goup, acute gangrenous group and the chronic group respectively, both in gradations of the appendix and the tube cavity expansion. Also there were statistically difference (P<0.05) between the acute gangrenous appendicitis group and the chronic group in the aspect of the appendix around effusion.ConclusionsUltrasound is valuable in the diagnosis of appendicitis, especially for acute gangrenous appendicitis and acute purulent appendicitis. The measurement of appendix with wide diameter, wall thickness by ultrasond preoperatively, and the characteristics of the wall layers, lumen expansion degree and the appendix around effusion are valuable in identifying chronic appendicitis, acute suppurative appendicitis, and acute gangrene appendicitis; The ultrasonic measurement of appendix wide diameter could identify acute gangrenous appendicitis and acute simple appendicitis. Ultrasound had limited value in identifying acute suppurative, acute gangrenous appendicitis, acute simple appendicitis, chronic appendicitis, and chronic appendicitis onset acute.

12.
Acta Universitatis Medicinalis Anhui ; (6): 1-4, 2014.
Article in Chinese | WPRIM | ID: wpr-445244

ABSTRACT

Objective To construct shRNA against optineurin ( OPTN ) clone and transfect the construction into HEK293FT cells for inhibiting the expression of OPTN, which is laid the foundation for further research of molecu-lar mechanism OPTN protein. Methods The insert of this clone was a double-strained DNA sequence against OPTN which would be transcripted into shRNA and it was synthesized according to the sequence in Origene. Ex-pression vector pRFP-C-RS was employed to fuse the insert to get the construction and finally confirmed by sequen-cing. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN and RFP signals could be detected using fluorescence microscope. Western blot was further employed to check the protein level of OPTN. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN was then obtained to investigate the functional study of OPTN which was measured by Salmonella infection experiment. Results shRNA eukaryotic expression vector targeting OPTN gene was successfully constructed. HEK293FT cells were transfected with pRFP-C-RS-shOPTN and 72 hours later we observed strong RFP signals showing that shOPTN expressed. Western blot was further employed to check the protein level of OPTN which showed that OPTN expression was indeed interfered. Salmonella infection assays was then performed and showed that OPTN could inhibit the proliferation of Salmonella that has invaded HEK293FT cells. Conclusion HEK293FT cell line which expresses shRNA against OPTN show a sharp inhibition of OPTN protein expression by 80%. So this cell line can be further used to investigate how OPTN regulates auto-phagy as an autophagy receptor. Meanwhile we find that OPTN can be as a autophagy regulator and strongly sup-press the proliferation of invasive Salmonella in vivo.

13.
Journal of Sun Yat-sen University(Medical Sciences) ; (6): 293-297, 2010.
Article in Chinese | WPRIM | ID: wpr-402763

ABSTRACT

[Objective]To evaluate the role of mature cumulus cells from oocyte-cumulus complex(OCC)in in-vitro maturation(IVM)and establish a new culture technique which is convenient to carry out.[Methods]The cumulus cells of OCC were cut off and dispersed by 1 mL syringe.The cumulus cells were co-cultured with the immature oocytes retrieved from the COH cycles after they adherent to the bottom of the dish.The immature oocytes were experienced IVM procedures in different culture media.They were divided into 3 groups(the oocytes at germinal vesicle stage from one woman were allotted to the same group randomly).Group 1(solution A):basic culture medium+human follicular fluid(hFF);Group 2(solution B):solution A+ cumulus cells(OCC);Group 3(solution C):solution A+ OCC+ follicle stimulating hormone(FSH)+ epidermal growth factor (EGF).Then,the maturation rate,fertilization rate and formation rate of available embryo were observed.[Results]In 113 treatment cycles,298 immature oocytes were performed IVM with solution A,B,and C.The difference for 24 hour maturation rates among 3 groups wag statistically significant(A:45.2%,B:61.7%,C:78.2%,P<0.05).There was no statistical difference for 25~48 hour maturation rates and normal fertilization rates of mature oocytes.The differences of cleavage rates and rescued embryo rates between group 1 and 2,group 1 and 3 were statistically significant(P<0.05).The formation rates of available embryo showed an increasing trend from group 1,2,to 3.[Conclusion]After being dispersed by simply beat upon with syringe and adherent culture,the mature cumulus eells from mature OCCs in COH cycles,together with growth factors in the follicular fluid or extraneously supplemented,could promote the IVM of immature oocyte.

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