1.Clinical study on the treatment of OSAHS in children with different surgical approaches based on H-uvu-lopalatopharyngoplasty
Zhitai CHEN ; Shengguo WANG ; Benzhong ZHOU ; Xianping SHI ; Feifeng WU ; Longqiao LI ; Zesheng ZHANG ; Guangjie ZHOU ; Yang LIU
Journal of Practical Stomatology 2024;40(5):687-690
Objective:To study the clinical effects of modified surgical method with H-uvulopalatopharyngoplasty(H-UPPP)and traditional surgical method in the treatment of obstructive sleep apnea hypopnea syndrome(OSAHS)in children.Methods:The clini-cal data of 364 pediatric patients with OSAHS were retrospectively analyzed.The patients were divided into 2 groups and treated by low-temperature plasma knife radio frequency ablation for bilateral tonsil removal combined with H-UPPP(group A,n=204)and low-temperature plasma knife radio frequency ablation only(group B,n=160)respectively.The operative time,intraoperative bleed-ing,postoperative secondary bleeding,VAS of pain following operation,postoperative traumatic white film shedding time,OAH1,LSaO2,ESS scores and complications of the 2 groups were compared.Results:In group A the operative time,intraoperative bleed-ing,VAS,OAHI and ESS scores were lower than those in group B(P<0.05),wile postoperative traumatic white film detachment time and LSaO2 were higher(P<0.05).No statistically significant difference in terms of the number of cases of postoperative seconda-ry bleeding and complications was observed between the 2 groups(P>0.05).Conclusion:The combination of low-temperature plas-ma radio frequency ablation and H-UPPP for the treatment of OSAHS may provide clear intraoperative view,and may improve the treatment effects.
2.Expression of Tim-3 in the peripheral blood of pancreatic cancer patients and the diagnostic value in combination with serum glucose chain antigen 19-9
Ke WANG ; Lijuan LIU ; Xianping CUI ; Wenying JIANG ; Changtao YU ; Jinxin SHI
Chinese Journal of Endocrine Surgery 2023;17(3):278-281
Objective:To investigate the expression of soluble T cell immunoglobulin and mucin domain-3 (Tim-3) in peripheral blood of patients with pancreatic cancer and its diagnostic value in combination with serum Carbohydrate antigen 19-9 (CA19-9) .Methods:106 newly diagnosed pancreatic cancer patients and 65 age and sex matched healthy individuals were enrolled. Tim-3 concentration was quantitatively determined by enzyme-linked immunosorbent assay (ELISA). According to the expression levels of soluble Tim-3 and serum CA19-9, a binary logistic regression model of receiver operating characteristic (ROC) curve was established to compare the diagnostic effects of serum CA19-9 and soluble Tim-3 alone or combined with the two tests.Results:The levels of soluble Tim-3 in the pancreatic cancer group were significantly higher than those in the healthy control group ( P<0.001). The expression level of soluble Tim-3 was significantly higher in patients with stage III-IV pancreatic cancer than in patients with stage I-II ( P=0.003). The AUC of soluble Tim-3 diagnosis for stage I-II pancreatic cancer was 0.856 (95%CI: 0.765 to 0.992 P<0.001), Serum CA19-9 The AUC used for the stage I-II pancreatic cancer diagnosis was 0.862 (95%CI: 0.772 to 0.926 P<0.001), The AUC for the combined diagnosis was 0.949 (95%CI: 0.880 - 0.985 P<0.001) ; In a healthy population and in patients with stage III-IV pancreatic cancer, the AUC of soluble T I I-IV pancreatic cancer in stage III was 0.927 (95%CI: 0.873 to 0.963 P<0.001), the AUC of serum CA19-9 used for the diagnosis of stage III-IV pancreatic cancer was 0.933 (95%CI: 0.881 to 0.968 P<0.001), the AUC for the combined diagnosis was 0.989 (95%CI: 0.956 to 0.999 P<0.001) . Conclusions:The combination of soluble Tim-3 and serum CA19-9 can improve the diagnostic rate of pancreatic cancer patients.
3. Impact and related mechanism on the improvement of hyperglycemia-induced pyroptosis in H9c2 cells by mircoRNA-214
Yan WANG ; Ranzun ZHAO ; Panke CHEN ; Guanxue XU ; Zhijiang LIU ; Xianping LONG ; Zhimei QIU ; Bei SHI
Chinese Journal of Cardiology 2019;47(10):820-828
Objective:
To investigate whether microRNA(miR)-214 can improve hyperglycemia induced pyroptosis in H9c2 cells through targeting caspase-1.
Methods:
H9c2 cells of rats those in good growth condition were selected and incubated into the T25 culture bottle after digestion and passage. Cells were cultured in an incubator at 37 ℃ with 5%CO2, repeat passage was made after cell density reached about 80%, The 5th to 8th generations of cells were selected for the subsequent experiments. To observe the effect of overexpression of miR-214 on pyroptosis and caspase-1 expression in H9c2 cells induced by hyperglycemia, the cells were divided into 4 groups: Control group(H9c2 cells cultured normally), Hyperglycemia group (HG group, 50 mmol/L glucose was used to intervene H9c2 cells for 24 hours), miR-214 mimics+hyperglycosis group (mimics+HG group, H9c2 cells were transfected with miR-214 mimics for 24 hours and then treated with 50 mmol/L hyperglycosis for 24 hours), miR-214 mimic-negative control+hyperglycaemic group(MNC+HG group, H9c2 cells were transfected with miR-214 mimic-negative control for 24 hours and then treated with 50 mmol/L hyperglycaemic for 24 hours). In order to further verify the anti-pyroptosis effect of miR-214 was mediated by targeted inhibition on caspase-1, cells overexpressing caspase-1 were used in the rescue experiment. The cells overexpressing caspase-1 were divided into 4 groups: Hyperglycemia group (HG group, 50 mmol/L glucose was used to intervene H9c2 cells for 24 hours), miR-214 mimics+hyperglycosis group (mimics+HG group, H9c2 cells were transfected with miR-214 mimics for 24 hours and then treated with 50 mmol/L hyperglycosis for 24 hours), miR-214 mimics+hyperglycosis+recombinant adenovirus (Ad-caspase-1-EGFP) group with caspase-1 gene and EGFP green fluorescent protein expression (mimics+HG+Ad-caspase-1-EGFP group, H9c2 cells were transfected with caspase-1-green fluorescent protein-carrying adenovirus for 48 hours, followed by transfection of miR-214 mimics for 24 hours, and then treated with 50 mmol/L hyperglycaemia for 24 hours), miR-214 mimics+HG+Ad-EGFP empty virus group (mimics+HG+Ad-EGFP group, H9c2 cells were transfected with empty adenovirus containing green fluorescent protein for 48 hours, followed by transfection with miR-214 mimics for 24 hours, and then treated with 50 mmol/L hyperglycosis for 24 hours). The mRNA expression levels of miRNA-214 and caspase-1 in cells were detected by real-time quantitative PCR. The expression and localization of caspase-1 protein were detected by immunofluorescence assay. Western blot was used to detect protein expression levels of procaspase-1, cleaved caspase-1, NLRP3 and ACS with β-actin as internal reference. The secretion of IL-1β and IL-18 in cell culture medium was detected by ELISA. The correlation between miR-214 and caspase-1 was detected by double luciferase reporter gene.
Results:
(1) The mRNA expression levels of miR-214 and caspase-1 in each group: the mRNA expressions of miR-214 in HG group and MNC+HG group were significantly lower than that in control group(
4.Impact and related mechanism on the improvement of hyperglycemia?induced pyroptosis in H9c2 cells by mircoRNA?214
Yan WANG ; Ranzun ZHAO ; Panke CHEN ; Guanxue XU ; Zhijiang LIU ; Xianping LONG ; Zhimei QIU ; Bei SHI
Chinese Journal of Cardiology 2019;47(10):820-828
Objective To investigate whether microRNA(miR)?214 can improve hyperglycemia induced pyroptosis in H9c2 cells through targeting caspase?1. Methods H9c2 cells of rats those in good growth condition were selected and incubated into the T25 culture bottle after digestion and passage. Cells were cultured in an incubator at 37℃with 5%CO2, repeat passage was made after cell density reached about 80%, The 5th to 8th generations of cells were selected for the subsequent experiments. To observe the effect of overexpression of miR?214 on pyroptosis and caspase?1 expression in H9c2 cells induced by hyperglycemia, the cells were divided into 4 groups: Control group(H9c2 cells cultured normally), Hyperglycemia group (HG group, 50 mmol/L glucose was used to intervene H9c2 cells for 24 hours), miR?214 mimics+hyperglycosis group (mimics+HG group, H9c2 cells were transfected with miR?214 mimics for 24 hours and then treated with 50 mmol/L hyperglycosis for 24 hours), miR?214 mimic?negative control+hyperglycaemic group(MNC+HG group, H9c2 cells were transfected with miR?214 mimic?negative control for 24 hours and then treated with 50 mmol/L hyperglycaemic for 24 hours). In order to further verify the anti?pyroptosis effect of miR?214 was mediated by targeted inhibition on caspase?1, cells overexpressing caspase?1 were used in the rescue experiment. The cells overexpressing caspase?1 were divided into 4 groups: Hyperglycemia group (HG group, 50 mmol/L glucose was used to intervene H9c2 cells for 24 hours), miR?214 mimics+hyperglycosis group (mimics+HG group, H9c2 cells were transfected with miR?214 mimics for 24 hours and then treated with 50 mmol/L hyperglycosis for 24 hours), miR?214 mimics+hyperglycosis+recombinant adenovirus (Ad?caspase?1?EGFP) group with caspase?1 gene and EGFP green fluorescent protein expression (mimics+HG + Ad?caspase?1?EGFP group, H9c2 cells were transfected with caspase?1?green fluorescent protein?carrying adenovirus for 48 hours, followed by transfection of miR?214 mimics for 24 hours, and then treated with 50 mmol/L hyperglycaemia for 24 hours), miR?214 mimics+HG+Ad?EGFP empty virus group (mimics+HG+Ad?EGFP group, H9c2 cells were transfected with empty adenovirus containing green fluorescent protein for 48 hours, followed by transfection with miR?214 mimics for 24 hours, and then treated with 50 mmol/L hyperglycosis for 24 hours). The mRNA expression levels of miRNA?214 and caspase?1 in cells were detected by real?time quantitative PCR. The expression and localization of caspase?1 protein were detected by immunofluorescence assay. Western blot was used to detect protein expression levels of procaspase?1, cleaved caspase?1, NLRP3 and ACS with β?actin as internal reference. The secretion of IL?1β and IL?18 in cell culture medium was detected by ELISA. The correlation between miR?214 and caspase?1 was detected by double luciferase reporter gene. Results (1) The mRNA expression levels of miR?214 and caspase?1 in each group: the mRNA expressions of miR?214 in HG group and MNC+HG group were significantly lower than that in control group(P<0.05). The mRNA expression of miR?214 in mimics+HG group was significantly higher than that in control group (P<0.05). The mRNA expression levels of caspase?1 in HG group and MNC+HG group were significantly higher than that in control group(P<0.05). The mRNA expression level of caspase?1 in mimics+HG group was lower than that in control group(P<0.05). (2) The expression of caspase?1 in each group: the green fluorescence intensity in the control group was weak, which was strong in the HG group and MNC+HG group. The green fluorescence expression was weaker in mimics+HG group than in HG group. (3) ASC and NLRP3 protein expression levels in each group: ASC and NLRP3 protein expression levels in HG group and MNC+HG group were higher than those in control group(P<0.05). ASC and NLRP3 protein expression levels were significantly lower in mimics+HG group than in mimics+HG group (P<0.05). (4) The secretion of IL?1β and IL?18 in the cell culture medium of each group: the content of IL?1β and IL?18 in the cell culture medium of HG group and MNC+HG group was significantly higher than that of control group (P<0.05). The content of IL?1β and IL?18 in the cell culture medium of mimics+HG group was significantly lower than that of the HG group (P<0.05). (5) Correlation between miR?214 and caspase?1: miR?214 specifically binds to caspase?1 3 'UTR. Meanwhile, Western blot results showed that cleaved caspase?1 protein expression levels were significantly higher in both HG group and MNC+HG group than in control group (P<0.05). The levels of cleaved caspase?1 were significantly lower in mimics+HG group than in HG group (P<0.05). There was no significant difference in procaspase?1 expression among groups (P>0.05). (6) The expression levels of procaspase?1, cleaved caspase?1, ASC and NLRP3 in each group in rescue experiment: there was no significant difference in the expression of procaspase?1 in each group (P>0.05). Cleaved caspase?1 , ASC and NLRP3 protein expressions were significantly lower in mimics+HG group than in HG group (P<0.05). However, cleaved caspase?1, ASC and NLRP3 protein expressions were significantly higher in mimics+HG+Ad?caspase?1?EGFP group than in mimics+HG group (P<0.05). (7) The expression of IL?1β and IL?18 in rescue experiment: the secretions of IL?1β and IL?18 in the cell culture medium of the mimics+HG group were significantly lower than that of HG group (P<0.05), which were significantly higher in mimics+HG+Ad?caspase?1?EGFP group than in mimics+HG group (P<0.05). Conclusion miR?214 can improve the hyperglycemia induced pyroptosis in H9c2 cells by targeted inhibition of the caspase?1.
5. Effect and related mechanism of miRNA-21 on hydrogen peroxide-induced C-kit+ cardiac stem cells apoptosis
Ranzun ZHAO ; Yan WANG ; Wenwen DENG ; Bei SHI ; Xianping LONG ; Zhenglong WANG ; Wenming CHEN
Chinese Journal of Cardiology 2017;45(11):971-977
Objective:
To explored the effect and related mechanism of miRNA-21 on hydrogen peroxide-induced C-kit+ cardiac stem cells apoptosis.
Methods:
C-kit+ cardiac stem cells were isolated from SD rats by the methods of enzyme digestion and magnetic bead. Cells were divided into the following experimental groups: (1) negative control mimics (NCM)group: cells were transfected with negative control miRNA-21 mimics for 48 hours; (2)mimics group: cells were transfected with miRNA-21 mimics for 48 hours; (3) NCM+ H2O2 group: negative control miRNA-21 mimics were transfected into cells for 48 hours and then treated with 100 μmol H2O2 for 2 hours; (4)mimics+ H2O2 group: miRNA-21 mimics were transfected into cells for 48 hours and then treated with 100 μmol H2O2 for 2 hours. mRNA of miRNA-21 was detected by RT-PCR. The apoptosis rate of C-kit+ cardiac stem cells was determined using the annexin V-FITC/PI staining assay. Western blot was employed to measure the expression of apoptosis related proteins(Caspase-3, Bax, and Bcl-2).
Results:
(1) Compared with the NCM group, the mRNA expression level of miRNA-21 was significantly up-regulated in mimics group, while obviously down-regulated in NCM+ H2O2 group(all
6.Clinical efficacy of comprehensive hyperbaric oxygen in treatment of the patients with sudden hearing loss
Zhitai CHEN ; Shengguo WANG ; Longqiao LI ; Xianping SHI ; Feifeng WU ; Houdong MEI ; Benzhong ZHOU
Chinese journal of nautical medicine and hyperbaric medicine 2017;24(6):478-481
Objective To investigate the clinical efficacy of comprehensive hyperbaric oxygen (HBO) in the treatment of the patients with sudden hearing loss .Methods Totally, 80 patients with sudden hearing loss who were admitted into our hospital for treatment from June 2015 to March 2017 were selected as research subjects, and were divided into the observation group (n =40) and the control group (n =40), in accordance with the sequence of admission.The control group was given volume expansion to improve hemorheology , vasodilator to improve microcirculation, glucocorticoids, neurotrophic drugs and ginkgo biloba extract dripping pills, while the observation group received HBO in addition to the treatment received by the control group .One course of treatment for all patients in the 2 groups was 10 days, and following 3 courses of treatment, changes in hearing and clinical efficacy before and after treatment for the patients of the 2 groups were compared between the 2 groups.Results Following treatment for 3 courses for the observation group , 4 cases were cured, 21 cases were markedly effective , 10 cases were effective and 5 cases were ineffective, with a total effective rate of 87.5%, while for the control group, no cases were cured, 5 cases were markedly effective, 22 cases were effective and 13 cases were ineffective, with a total effective rate of 67.5%.Statistical significance could be seen, when comparisons were made between the observation group and the control group (P <0.05).Before treatment, there was no significant differences in full frequency hearing loss for the patients of 2 groups (P >0.05).After 2 courses of treatment, the value of low frequency hearing for the observation group increased more significantly, as compared with that of the control group [(37.39 ±5.77) vs (23.91 ±4.08) dB,P <0.05], however, there were no statistical differences in the increased values of high frequency hearing , when comparisons were made between the 2 groups (P >0.05).After 3 courses of treatment, the increased values of both low frequency and high frequency hearing for the patients of the observation group were all significantly better than those of the control group [ low frequency: (45.99 ±5.98) vs (36.79 ±5.72) dB; high frequency:(19.63 ±3.94) vs (9.70 ±2.17)dB, P <0.05].Conclusions Comprehensive HBO treatment could effectively improve the hearing level of the patients with full frequency sudden hearing loss , which was worth further clinical popularization .
7.Clinical efficacy of comprehensive hyperbaric oxygen in treatment of the patients with sudden hearing loss
Zhitai CHEN ; Shengguo WANG ; Longqiao LI ; Xianping SHI ; Feifeng WU ; Houdong MEI ; Benzhong ZHOU
Chinese journal of nautical medicine and hyperbaric medicine 2017;24(6):478-481
Objective To investigate the clinical efficacy of comprehensive hyperbaric oxygen (HBO) in the treatment of the patients with sudden hearing loss .Methods Totally, 80 patients with sudden hearing loss who were admitted into our hospital for treatment from June 2015 to March 2017 were selected as research subjects, and were divided into the observation group (n =40) and the control group (n =40), in accordance with the sequence of admission.The control group was given volume expansion to improve hemorheology , vasodilator to improve microcirculation, glucocorticoids, neurotrophic drugs and ginkgo biloba extract dripping pills, while the observation group received HBO in addition to the treatment received by the control group .One course of treatment for all patients in the 2 groups was 10 days, and following 3 courses of treatment, changes in hearing and clinical efficacy before and after treatment for the patients of the 2 groups were compared between the 2 groups.Results Following treatment for 3 courses for the observation group , 4 cases were cured, 21 cases were markedly effective , 10 cases were effective and 5 cases were ineffective, with a total effective rate of 87.5%, while for the control group, no cases were cured, 5 cases were markedly effective, 22 cases were effective and 13 cases were ineffective, with a total effective rate of 67.5%.Statistical significance could be seen, when comparisons were made between the observation group and the control group (P <0.05).Before treatment, there was no significant differences in full frequency hearing loss for the patients of 2 groups (P >0.05).After 2 courses of treatment, the value of low frequency hearing for the observation group increased more significantly, as compared with that of the control group [(37.39 ±5.77) vs (23.91 ±4.08) dB,P <0.05], however, there were no statistical differences in the increased values of high frequency hearing , when comparisons were made between the 2 groups (P >0.05).After 3 courses of treatment, the increased values of both low frequency and high frequency hearing for the patients of the observation group were all significantly better than those of the control group [ low frequency: (45.99 ±5.98) vs (36.79 ±5.72) dB; high frequency:(19.63 ±3.94) vs (9.70 ±2.17)dB, P <0.05].Conclusions Comprehensive HBO treatment could effectively improve the hearing level of the patients with full frequency sudden hearing loss , which was worth further clinical popularization .
8.Transfection of calcitonin gene-related peptide mediated by lentivirus vec-tor in vitro and its effects on cardiac stem cell viability
Jidong RONG ; Ling LI ; Xianping LONG ; Wenwen DENG ; Bei SHI
Chinese Journal of Pathophysiology 2016;32(8):1445-1449,1456
AIM:To investigate the effect of calcitonin gene-related peptide (CGRP)transfection into c-kitpos cardiac stem cells (c-kit+CSCs) on the cell viability.METHODS: Under the sterile condition, the auricles of SD rats were taken out , and then c-kit +CSCs were collected through enzyme digestion and immunomagnetic bead separation (MACS).The cells were identified by flow cytometry .c-kit +CSCs were transfected with enhanced green fluorescent pro-tein CGRP lentiviral vector ( Lv-EGFP-CGRP) or enhanced green fluorescent protein lentiviral vector ( Lv-EGFP).The cells were randomly divided into Lv-EGFP-CGRP-CSCs group , Lv-EGFP-CSCs group and CSCs group .The transfection was observed under the fluorescence microscope .The transfection efficiency was detected by flow cytometry .The CGRP protein secretion in the cell culture supernatants was detected by ELISA .The viability of c-kit+CSCs transfected with Lv-EGFP-CGRP or Lv-EGFP was measured by CCK-8 assay.RESULTS:c-kit+CSCs were isolated and cultured successfully .The expression positive rate of c-kit was 91.0%and the expression positive rates of CD 45 and CD34 were 4.5% and 4.0%, respectively.After transfected with lentivirus for 48 h, the stable fluorescence in c-kit +CSCs was observed under fluores-cence microscope .The transfection efficiency were 80%when MOI was 20.The level of CGRP was significantly increased in Lv-ECFP-CGRP-CSCs group compared with Lv-EGFP-CSCs group and CSCs group (P<0.05).Meanwhile, transfection with lentiviral vector in each group did not affect the viability of c-kit+CSCs.CONCLUSION:Transfection of Lv-EGFP-CGRP into c-kit+CSCs was successful .The secretion of CGRP was found in the transfected c-kit+CSCs and the viability was not changed after transfection .CGRP-modified c-kit+CSCs may play a role in treating myocardial infarction .
9.Observation for the Effect of Vacuum Sealing Drainage in Treating the Patients With Pacemaker Pocket Infection
Dongmei WANG ; Bei SHI ; Xianping LONG ; Jidong RONG
Chinese Circulation Journal 2016;31(10):1006-1008
Objective: Pocket infection in patients after total removal of implanted pacemaker has many problems for their electronic system; our research intends to explore the feasibility of conservatively treating such infection and retain the electronic system. Methods: A total of 4 patients with pacemaker pocket infection in our hospital from 2015-01 to 2016-02 were studied. Thorough debridement and disinfection were conducted in infected pockets and devices, meanwhile vacuum sealing drainage was applied. Electrode wire was kept and intravenous antibiotics were given for (7-10) days after the operation in all patients. Results: The average time of infection occurred at 14.75 months after operation with the type of isolated pacemaker pocket infection. Pocket vacuum suction drainage was performed, with the mean of 7.25 (5-10) months follow-up observation, infection was disappeared and the patients had good wound healing. Conclusion: With thorough debridement of infected pocket, rational treatment of residual electronic system and vacuum sealing drainage, the infection might be effectively controlled for complete recovery without lead extraction in relevant patients.
10.Lentiviral vector mediated CGRP gene in vitro transfection and its effects on biological properties of MSC
Panke CHEN ; Bei SHI ; Guanxue XU ; Zhijiang LIU ; Xianping LONG ; Wei ZHANG ; Shuai MA
Chongqing Medicine 2015;(14):1873-1875,1878
Objective To explore in vitro mesenchymal stem cell (MSC) transfection of lentiviral vector mediated calcitonin gene‐related peptide(CGRP) gene and its effects on biological properties of MSC .Methods MSC were isolated ,cultured and identi‐fied .MSC were infected by lentivirus encoding recombinant enhanced green fluorescent protein (EGFP) gene and CGRP (Lv‐EG‐FP‐CGRP) .The transfection efficiency was determined by the inverted fluorescence microscope and flow cytometry .The expression levels of CGRP were detected in CGRP‐modified MSC by using real‐time PCR ,immunocytochemistry and enzyme‐linked immu‐nosorbent assay (ELISA) .The proliferation ,aging and differentiation ability of MSC were evaluated by MTT ,β‐galactosidase stai‐ning and inducing differentiation respectively .Results After 48 h of MSC transfection by Lv‐EGFP‐CGRP ,EGFP/CGRP could be expressed stably .When multiplicity of infection (MOI) was 30 ,the transfection efficiency reached more than 80% .Compared with the MSC group and the MSC‐EGFP group ,the mRNA and protein expression levels of CGRP in CGRP‐modified MSC(MSC‐CGRP group) were markedly increased(all P<0 .01) .The results of MTT ,β‐galactosidase staining and inducing differentiation assay dem‐onstrated that the transfected CGRP basically had no effect on the proliferation ,aging and endotheliocyte differerntiation of MSC . Conclusion MSC is a kind of ideal genetic vector cell ,which can serve as the target cell of CGRP gene transduction for the applica‐tion of gene therapy and lays the foundation for follow‐up in vitro and vivo experiments .

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