1.Expression and subcellular localization of urocortin in syncytiotrophoblast of human term placenta
Yingyuan ZHU ; Zineng WANG ; Yike ZENG ; Peie ZHENG ; Jianping XU ; Zuwen GUO ; Fuxing TANG
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine.
2.Application of diffusion weighted imaging in accurate diagnosis of middle ear cholesteatoma
Fang GUO ; Dejun ZHANG ; Zeming FU ; Yingyuan GUO ; Yining WAN ; Guofang GUAN ; Junfeng LYU ; Xiuying CHEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2020;55(11):1037-1042
Objective:To evaluate the accuracy of diffusion weighted magnetic resonance imaging (DWI-MRI) combined with high resolution temporal bone CT (HRCT) in the location diagnosis of middle ear cholesteatoma and its value in the postoperative follow-up.Methods:134 patients with inital cholesteatoma and 22 patients with suspected recurrent cholesteatoma were selected for HRCT, conventional MRI and DWI examination. Based on the intraoperative and pathological diagnosis, DWI and HRCT images were combined to evaluate the consistency between the lesion location and invasion area of the initial cholesteatoma and intraoperative lesions. The results of HRCT and DWI in the diagnosis of recurrent middle ear cholesteatoma were statistically analyzed to evaluate their diagnostic efficacy.Results:The accuracy rate of DWI combined with HRCT was 90.3%.The sensitivity, specificity, positive predictive value and negative predictive value of HRCT and DWI in the diagnosis of recurrent middle ear cholesteatoma were 27.8%, 75.0%, 83.3%, 18.8% and 100%, 75.0%, 94.7% and 100%, respectively, and the Kappa values consistent with the pathological results were 0.024 and 0.843, respectively. Chi-square test confirmed that there were differences in the diagnosis between groups ( P<0.001). Conclusions:Combined with the high sensitivity of DWI and the high resolution of HRCT, the accuracy of preoperative positioning of the newly diagnosed cholesteatoma can be improved and surgery strategy can be guided. DWI is also of high diagnostic value for recurrent cholesteatoma in the middle ear.
3.Changes in circadian gene cryptochrome 2 expression in mouse models of psoriasis and HaCaT cells and their underlying mechanisms
Lingling YAO ; Zengyang YU ; Chunyuan GUO ; Jing ZHOU ; Lian CUI ; Qian YU ; Yingyuan YU ; Xue ZHOU ; Jiangluyi CAI ; Yuling SHI
Chinese Journal of Dermatology 2022;55(9):759-766
Objective:To investigate changes in circadian gene cryptochrome 2 (CRY2) expression in mouse models of psoriasis and HaCaT cells, and to explore underlying mechanisms.Methods:Imiquimod-induced mouse model experiment: 12 C57BL/6 female mice were randomly and equally divided into imiquimod group receiving topical imiquimod treatment for 5 consecutive days and control group receiving no treatment; these mice were sacrificed on day 6, skin tissues were resected from the back of mice, and immunofluorescence staining was performed to determine the CRY2 expression in the epidermis. HaCaT cell transfection experiment: HaCaT cells with small interfering RNA (siRNA) -mediated knockdown of CRY2 served as siRNA-CRY2 group, and siRNA-NC group as control group; 5-ethynyl-2′-deoxyuridine (EdU) staining was performed to evaluate the proliferative activity of the HaCaT cells, real-time fluorescence-based quantitative PCR (qPCR) to determine the mRNA expression of chemokines in the HaCaT cells, and Western blot analysis to determine phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2) . Tumor necrosis factor-α (TNF-α) -stimulated animal and cell experiments: 12 C57BL/6 female mice were randomly and equally divided into TNF-α group subcutaneously injected with TNF-α solution in the ear for 6 days, and phosphate buffered saline (PBS) group subcutaneously injected with the same amount of PBS; the mice were sacrificed on day 7, skin tissues were resected from the ear of mice, and immunofluorescence staining was conducted to determine the CRY2 expression in the epidermis; CRY2-knockdown HaCaT cells stimulated with 50 ng/ml TNF-α for 12 hours served as siRNA-CRY2 + TNF-α group, and siRNA-NC + TNF-α group as control group; qPCR was performed to determine the mRNA expression of chemokines in HaCaT cells in the above groups. Statistical analysis was carried out by using two-independent-sample t test. Results:Immunofluorescence staining showed that the CRY2 protein expression was significantly lower in the mouse dorsal epidermis in the imiquimod group (0.94 ± 0.23) than in the control group (2.30 ± 0.25, t = 3.99, P = 0.016) . Compared with the siRNA-NC group, the siRNA-CRY2 group showed significantly increased proportions of EdU-positive cells (48.13% ± 10.97% vs. 38.23% ± 0.81%, t = 5.00, P = 0.007) , mRNA expression levels of chemokines CXCL1 and CXCL8, as well as significantly increased phosphorylated (p) -ERK1/2 protein expression levels (all P < 0.05) , while there were no significant differences in the CCL20 mRNA expression or ERK1/2 protein expression between the two groups (both P > 0.05) . Immunofluorescence staining showed significantly decreased CRY2 protein expression level in the mouse ear epidermis in the TNF-α group (0.37 ± 0.34) compared with the PBS group (2.04 ± 0.17, t = 4.38, P = 0.012) ; the relative mRNA expression levels of chemokines CXCL1, CXCL8, and CCL20 in HaCaT cells were significantly higher in the siRNA-CRY2 + TNF-α group than in the siRNA-NC + TNF-α group (all P < 0.05) . Conclusion:CRY2 was markedly underexpressed in psoriasis, which might promote the proliferation of keratinocytes and expression of chemokines CXCL1, CXCL8 and CCL20, and TNF-α might be an upstream cytokine that could downregulate CRY2 expression.
4.Research progress on the crown-implant ratio and clinical complications
XU Shulan ; GUO Zehong ; NING Yingyuan ; GAO Yan
Journal of Prevention and Treatment for Stomatological Diseases 2020;28(9):545-550
The crown-root ratio (C/R) theory of natural teeth has been widely recognized in the field of stomatology,and has important clinical significance in predicting and assessing the prognosis of natural teeth as well as for abutment selection during denture restoration. In the past few decades, scholars have advocated for the implantation of implants as long in length as possible to improve the success rate according to the theory of crown-root ratio of natural teeth. However, with the application of short implants, our philosophy of implantation has changed, and the relationship between the crown-implant (C/I) ratio and complications has become one of the current research hotspots. In this paper, the concept of the crown-implant ratio, the research progress of the C/I ratio, the implant survival rate and clinical complications of implant restoration were reviewed and summarized, and the following suggestions were put forward: although most studies have shown no significant correlation between the C/I ratio and implant survival or marginal bone loss, this relationship may increase the risk of mechanical complications. A C/I < 3 and a crown length < 15 mm are recommended in implant restoration; when ultra-short implants are applied, the implant system can increase the bone-to-implant contact area, and splint prostheses such as crown or bridge are recommended.
5.Clinical observation of alveolar ridge approach for odontogenic maxillary sinusitis and implant restoration
GUO Zehong ; NING Yingyuan ; XU Shulan ; ZHAN jieling ; DING Xianglong ; GAO Yan
Journal of Prevention and Treatment for Stomatological Diseases 2019;27(8):505-509
Objective :
To explore the clinical effect of the alveolar crest approach in the treatment of odontogenic maxillary sinusitis and the repair of edentulous implants in this area.
Methods:
This was a retrospective case series of 20 patients with odontogenic sinusitis. The pathogenesis in each case was investigated. After elimination of the dental origin, each patient was treated with flushing, drainage and anti-inflammatories through the alveolar crest approach. Postoperative CBCT reexamination was performed to evaluate the therapeutic effect. Maxillary sinus elevation surgery with simultaneous or delayed implantation was performed after maxillary sinusitis healing was confirmed. The patients were followed postoperatively.
Results :
Twenty patients with odontogenic sinusitis were treated by the alveolar crest approach, and 17 were cured, for a cure rate of 85%. Among them, 17 of the maxillary sinusitis patients were followed for 1 year, with good results.
Conclusion
The alveolar crest approach is feasible for the treatment of odontogenic maxillary sinusitis and can serve as a minimally invasive method for the repair of edentulism in this area and implantation.
6.Effect of laser-etched pure titanium surface on early proliferation of MG63 cells
GUO Zehong ; NING Yingyuan ; XU Shulan ; ZHU Peijun ; DING Xianglong ; GAO Yan
Journal of Prevention and Treatment for Stomatological Diseases 2019;27(7):435-440
Objective :
To investigate the effect of a laser-etched pure titanium surface on proliferation of the human osteosarcoma cell line MG63 and to provide a basis for study of implant surface modification.
Methods:
The pure titanium plate was cut into titanium pieces by a numerical control machine tool and divided into smooth surface and laser etching groups. The titanium surface of the laser etching group was etched with an Nd:YAG continuous wave laser using predetermined parameters, and the surfaces were observed by scanning electron microscopy (SEM). The surface micromorphology of each titanium sheet was evaluated. The relative element content of the titanium surface was measured by energy dispersive X-ray spectroscopy (EDS). The Ra value of each surface was determined using the Veeco roughness tester. MG63 cells were inoculated on 2 sets of titanium tablets. At 1, 3, and 6 h postinoculation, cell adhesion to the two groups of titanium sheets was observed under the microscope. At 24 h after inoculation, cellular F-actin was directly stained using immunofluorescence, and the morphology of the cytoskeleton was observed by laser confocal microscopy. Cell proliferation was examined at 1, 3, and 5 d using a MTS kit, and the data were analyzed with SAS 9.4.
Results :
The surface of the smooth surface group was smooth and flat, the element composition was pure titanium, and the roughness Ra was 179.23 nm. The surface of the laser-etched group formed a regular and uniform pore structure. The composition was mainly Ti, O, C, etc, and the surface roughness Ra was 14.11 μm. A large number of cells were uniformly distributed on the two titanium sheets in the observations at 1, 3, and 6 h. At 24 h postinoculation, MG63 cells were completely stretched on the two sets of titanium sheets and had extended a large number of pseudopods and microfilaments to cross-link with peripheral cells; moreover, the cell division phase was observed. The cell proliferation of the two groups at 1, 3, and 5 d showed a significant increase with time, indicating that no cytotoxicity occurred on the surfaces of the two groups. However, the cell proliferation in the laser-etched group was superior to that in the mechanical smooth surface group.
Conclusion
The surface morphology of titanium can be controlled by laser etching, which is conductive to increase the microstructure of implants without cytotoxicity and promoting osteoblast proliferation in the early stage.
7.Comparing the prognostic value of 3 diagnostic criteria of bronchopulmonary dysplasia in preterm infants
Xin WANG ; Jing GUO ; Yanyan WU ; Yangke LU ; Dapeng LIU ; Mingchao LI ; Rui LI ; Yingyuan WANG ; Wenqing KANG
Chinese Journal of Pediatrics 2024;62(1):36-42
Objective:To compare the prognostic value of 3 diagnostic criteria of bronchopulmonary dysplasia (BPD) in preterm infants with gestational age<32 weeks.Methods:The retrospective cohort study was conducted to collect the clinical data of 285 preterm infants with BPD admitted to the Department of Neonatology, Children′s Hospital Affiliated to Zhengzhou University from January 2019 to September 2021, who were followed up regularly after discharge. The primary composite adverse outcome was defined as death or severe respiratory morbidity from 36 weeks of corrected gestational age to 18 months of corrected age, and the secondary composite adverse outcome was defined as death or neurodevelopmental impairment. According to the primary or secondary composite adverse outcomes, the preterm infants were divided into the adverse prognosis group and the non-adverse prognosis group. The 2001 National Institute of Child Health and Human Development (NICHD) criteria, 2018 NICHD criteria, and 2019 Neonatal Research Network (NRN) criteria were used to diagnose and grade BPD in preterm infants. Chi-square test, Logistic regression analysis, receiver operating characteristic (ROC) curve and Delong test were used to analyze the prognostic value of the 3 diagnostic criteria.Results:The 285 preterm infants had a gestational age of 29.4 (28.1, 30.6) weeks and birth weight of 1 230 (1 000, 1 465) g, including 167 males (58.6%). Among 285 premature infants who completed follow-up, the primary composite adverse outcome occurred in 124 preterm infants (43.5%), and the secondary composite adverse outcome occurred in 40 preterm infants (14.0%). Multivariate Logistic regression analysis showed that severe BPD according to the 2001 NICHD criteria, gradeⅡand Ⅲ BPD according to the 2018 NICHD criteria and grade 2 and 3 BPD according to the 2019 NRN criteria were all risk factors for primary composite adverse outcomes (all P<0.05). ROC curve showed that the area under the curve (AUC) of the 2018 NICHD criteria and 2019 NRN criteria were both higher than that of the 2001 NICHD criteria (0.70 and 0.70 vs. 0.61, Z=4.49 and 3.35, both P<0.001), but there was no significant difference between the 2018 NICHD and 2019 NRN criteria ( Z=0.38, P=0.702). Multivariate Logistic regression analysis showed that the secondary composite adverse outcomes were all associated with grade Ⅲ BPD according to the 2018 NICHD criteria and grade 3 BPD according to the 2019 NRN criteria (both P<0.05). ROC curve showed that the AUC of the 2018 NICHD criteria and 2019 NRN criteria were both higher than that of the 2001 NICHD criteria (0.71 and 0.71 vs. 0.58, Z=2.93 and 3.67, both P<0.001), but there was no statistically significant difference between the 2018 NICHD and 2019 NRN criteria ( Z=0.02, P=0.984). Conclusion:The 2018 NICHD and 2019 NRN criteria demonstrate good and comparable predictive value for the primary and secondary composite adverse outcomes in preterm infants with BPD, surpassing the predictive efficacy of the 2001 NICHD criteria.