[The mechanism of NRF2 inhibiting ROS induced autophagy to reduce ovarian granulosa cells damage].

This study explores the effects and possible mechanisms of nuclear factor E2 related factor 2 (NRF2) on ovarian granulosa cells, providing a scientific basis to prevent premature ovarian failure. An ovarian cell injury model was constructed by treating human ovarian granulosa cell (KGN cell) with 4-Vinylcyclohexene dioxide (VCD). Firstly, KGN cells were treated with different concentrations of VCD, and cell counting kit 8 (CCK-8) was used to detect ovarian cell proliferation. After determining IC50 by CCK8, the levels of estradiol and progesterone in the cell supernatant were detected using enzyme-linked immunosorbent assay (ELISA), reactive oxygen species (ROS) assay kit was used to detect the content of ROS in ovarian cells, real-time fluorescence quantitative polymerase chain reaction (qRT PCR) was used to detect the mRNA expression level of NRF2, and Western blot was used to detect the protein expression level of NRF2. Further, NRF2 silence (siNRF2) and overexpression (NRF2-OE) cell models were constructed through lentivirus transfection, and the effects of regulating NRF2 on VCD treated cell models were investigated by detecting hormone levels, oxidative stress indicators (ROS, SOD, GSH-Px), and autophagy (LC3B level). The results showed that VCD intervention inhibited the proliferation of ovarian granulosa cells in a time-dependent and dose-dependent manner (F>100, P<0.05), with an IC50 of 1.2 mmol/L at 24 hours. After VCD treatment, the level of estradiol in the cell supernatant decreased from (56.32±10.18) ng/ml to (24.59±8.75) ng/ml (t=5.78, P<0.05). Progesterone decreased from (50.25±7.03) ng/ml to (25.13±6.67) ng/ml (t=6.54, P<0.05). After VCD treatment, the SOD of cells decreased from (44.47±7.71) ng/ml to (30.92±4.97) ng/ml (t=3.61, P<0.05). GSH-Px decreased from (68.51±10.17) ng/ml to (35.19±6.59) ng/ml (t=5.73, P<0.05). Simultaneously accompanied by an increase in autophagy and a decrease in NRF2. This study successfully constructed KGN cell models that silenced NRF2 and overexpressed NRF2. Subsequently, this study treated each group of cells with VCD and found that the cell proliferation activity of the siNRF2 group was significantly reduced (t=8.37, P<0.05), while NRF2-OE could reverse the cell activity damage caused by VCD (t=3.37, P<0.05). The siNRF2 group had the lowest level of estradiol (t=5.78, P<0.05), while NRF2-OE could reverse the decrease in cellular estradiol levels caused by VCD (t=5.58, P<0.05). The siNRF2 group had the lowest progesterone levels (t=3.02, P<0.05), while NRF2-OE could reverse the decrease in cellular progesterone levels caused by VCD (t=2.41, P<0.05). The ROS level in the siNRF2 group was the highest (t=2.86, P<0.05), NRF2-OE could reverse the increase in ROS caused by VCD (t=3.14, P<0.05), the SOD enzyme content in the siNRF2 group was the lowest (t=2.98, P<0.05), and NRF2-OE could reverse the decrease in SOD enzyme content caused by VCD (t=4.72, P<0.05). The GSH-Px enzyme content in the siNRF2 group was the lowest (t=3.67, P<0.05), and NRF2-OE could reverse the decrease in antioxidant enzyme content caused by VCD (t=2.71, P<0.05). The LC3B level was highest in the siNRF2 group (t=2.45, P<0.05), and NRF2-OE was able to reverse the LC3B elevation caused by VCD (t=9.64, P<0.05). In conclusion, NRF2 inhibits ROS induced autophagy, thereby playing a role in reducing ovarian granulosa cell damage, which may be a potential target for premature ovarian failure.

[Clinicopathological characteristics of Klinefelter syndrome: a testicular biopsy analysis of 87 cases].

Objective: To investigate the clinicopathological characteristics of testicular biopsies from Klinefelter syndrome (KS) patients. Methods: The testicular biopsy specimens of 87 patients with KS (a total of 107 biopsy specimens) were collected from the Department of Pathology, Peking University Third Hospital, Beijing, China from January 2017 to July 2022. All patients were diagnosed as KS by peripheral blood karyotyping analysis. The testicular histopathologic features, testicular volume and hormone levels were evaluated retrospectively. The histopathologic analysis was used to assess the quantity and morphology of Leydig cells, the spermatogenic state of seminiferous tubules, the thickening of the basement membrane of seminiferous tubules and the changes of stroma. Results: Leydig cell proliferative nodules were seen in 95.3% (102/107) of KS testicular biopsy tissues. The eosinophilic inclusion bodies and lipofuscin in Leydig cells were found in 52.3% (56/107) and 57.9% (62/107) of specimens, respectively. The Sertoli cell only seminiferous tubules and the hyalinized tubules were found in 66.4% (71/107) and 76.6% (82/107) of the examined tissues, respectively. The tubules with complete spermatogenic arrest were found in 15.9% (17/107) of specimens, and 5.6% (6/107) of the specimens showed low spermatogenesis or incomplete spermatogenic arrest. In 85.0% (91/107) of the specimens, increased thick-walled small vessels with hyaline degeneration were identified. Conclusions: The most common features of KS testicular specimens are Leydig cell proliferative nodules, hyaline degeneration of seminiferous tubules and proliferation of thick-walled blood vessels. Testicular biopsy specimens of KS are rare. The pathologists can make a tentative diagnosis of KS based on the histological findings, combined with the ultrasound and laboratory results, which is helpful for further diagnosis and treatment of KS.

Silencing of linc00337 inhibits proliferation, cell cycle progression, migration, and invasion of colorectal cancer cells through the MEK/ERK pathway.

OBJECTIVE: To explore the expression and biological functions of linc00337 in colorectal cancer (CRC), as well as its underlying mechanism. PATIENTS AND METHODS: The relative expression of linc00337 in 47 cases of CRC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The si-linc00337 interference sequences were designed and transiently transfected into CRC cells. The interference efficiency was detected via qRT-PCR. Regulatory effect of linc00337 on proliferation of CRC cells was detected via colony formation assay. Cell cycle distribution and apoptosis rate after interference in linc00337 expression were determined using flow cytometry. Moreover, the effects of linc00337 knockdown on cell migration and invasion were detected using transwell assay. At last, the effect of si-linc00337 on the MEK/ERK signaling pathway was detected using Western blotting. RESULTS: The results of qRT-PCR showed that among the 47 cases of CRC tissues, the expression of linc00337 was up-regulated in 40 cases. Similarly, it was highly expressed in CRC cell lines. The results of colony formation assay manifested that cell proliferation declined after interference in linc00337 expression. The results of flow cytometry and transwell assay showed that interference in linc00337 expression arrested the cell cycle in G1/G0 phase, increased the apoptosis rate, and inhibited the invasion and migration of CRC cells. According to the results of Western blotting, expressions of molecular markers in the MEK/ERK pathway after interference in linc00337 expression were significantly changed. CONCLUSIONS: Linc00337 is up-regulated in CRC tissues and cells. Interference in linc00337 expression can inhibit cell proliferation, migration, and invasion and promote apoptosis through the MEK/ERK pathway.

[Demographic characteristics and canalith repositioning efficacy in 907 patients with typical benign paroxysmal positional vertigo].

AbstractObjective:To review the demographic characteristics and canalith repositioning efficacy in 907 patients with typical benign paroxysmal positional vertigo(BPPV). Method: The demographic characteristics of 907 patients with typical BPPV were statistically analyzed. According to the type of BPPV, patients were treated with the appropriate repositioning maneuver, and the clinical efficacy of repositioning maneuver was analyzed and summarized. Result: Nine hundred and seven patients of BPPV with typical nystagmus were elected in this study. 585 out of 907 were female and 322 out of were male, and the mean age was 53.10±14.25(13 to 89) years. The lesion located to the posterior semiCIrcular canal was 489 patients(53.9%), horizontal semiCIrcular canal was 312 patients(34.4%), anterior semiCIrcular was 63 patients(6.9%), and multiple semiCIrcular was 43 patients(4.8%). According to Kaplan-Meier survival analysis curve, the median cure time for the modified Semont and Epley repositioning maneuver in PC-BPPV groups was 3 days, and there was no significant difference in survival curves between the two repositioning maneuver. Meanwhile, the median cure time for Barbecue and Li horizontal canal quick repositioning maneuver groups was 3 days, and 1 day for Gufoni repositioning maneuver group in HC-BPPV groups. And there was no significant difference in survival curves among the three repositioning maneuver. In the AC-BPPV, 7 cases, 31 cases, 57 cases, 58 cases were cured with the Li anterior canal quick repositioning maneuver in the 1st day, the 3rd day, 1 week later, and 1 month later, and 5 cases lost to be follow-up. According to Kaplan-Meier survival analysis curve, the median cure time for the Li anterior canal quick repositioning maneuver in AC-BPPV groups was 3 days. Conclusion:Repositioning maneuver represents a simple, safe, rapid and effective approach to the treatment of BPPV. Therefore, repositioning maneuver should be choice for the BPPV treatment.

[Treatment of patients with probable benign paroxysmal positional vertigot].

Objective:To study the treatment and outcome of the patients with probable BPPV who lacked typical nystagmus.Method： Nine hundred and twenty-two patients with posterior canal BPPV or lateral canal BPPV were selected from the database of our department. There were 801 patients with typical nystagmus during positional testing and 121 patients lacked of typical nystagmus. Manual reposition (Eply,modified Semount,Barbecue and Li manuve) were used and the follow-up was（3.7±2.5）days.Result：In the patients with probable BPPV,there were 44 patients affected by posterior canal and 77 patients had been affected by lateral canal.In the patients with definite BPPV,there were 489 patients affected by posterior canal and 312 by horizontal canal.There was no statistic difference in the result of the treatment (cure,relief and no response) between the probable BPPV and definite BPPV (P>0.05).However,the rate of 'cure' and 'relief' in the probable BPPV was higher than the definite BPPV in lateral canal (P=0.023).Conclusion:A few patients with probable BPPV may develop to definite BPPV or other disease with dizziness.Maneuver of reposition is meaningful for probable BPPV,especially for patients with lateral canal probable BPPV.

[Impact of apical preparation diameter on fracture resistance of mandibular premolar roots].

OBJECTIVE: To compare the fracture resistance of roots of mandibular premolar with different apical preparation diameters. METHODS: Sixty single-rooted single canal permanent mandibular premolar teeth extracted newly for orthodontic reason without immatureness, fracture or cracks were selected, with a curvature less than 10°, and internal length: short diameter of less than 2 at a level 5 mm from the apex. All the teeth were decoronated, leaving roots 13 mm in length. The initial apical file size for the teeth was ≤15#. The roots were assigned to 6 groups based on weights with random block design. Group A: blank control group, no instrumentation was performed. Groups B-F: the master apical file (MAF) was 40#, 45#, 50#, 55# and 60#, respectively. In the five experimental groups the roots were instrumented using hand files with step-back technique at 1 mm increments, resulting in a taper of 0.05. The irrigant used was distilled water. After mounted in acrylic resin, all the teeth were subject to vertical loading using an Instron testing machine until fractured. The occurrence of fractures was detected when the applied load suddenly decreased. The fracture load values and fracture modes were recorded. One-way ANOVA and post-hoc Tukey test were used to determine the difference of fracture load values between the groups (P<0.05). Chi-square tests were used to compare the modes of root fracture. RESULTS: Five experimental groups exhibited lower fracture load values than that of control group [(1 444±155) N]. The mean fracture load values for roots instrumented to an apical diameter of 50# [(1 027±128) N], 55# [(994±150) N] and 60# [(983±166) N] were significantly lower than that of control group and 40# group [(1 339±131) N] and 45# [(1 287±144) N] (P<0.05). Buccal-lingual fracture, mesio-distal fracture and compound fracture occurred 55%, 13% and 32%, respectively. No difference of fracture mode was detected in the six groups. CONCLUSION: The fracture resistance reduced significantly when the roots were instrumented to an apical diameter of 50# or larger.

[A randomized controlled trial on short-term efficacy of the modified Semont maneuver for the treatment of posterior canal benign paroxysmal positional vertigo].

Objective:To observe the short-term efficacy of modified Semont maneuver for posterior canal benign paroxysmal positional vertigo (PC-BPPV)Method:This was a prospective randomized controlled trial on 130 PC-BPPV patients. Subjects were randomized divided into two groups: modified Semont (65 patients),and Epley(65 patients).Each maneuver was repeated twice,and the presence of sequelae,nystagmus and vertigo on positional testing were evaluated 3rd day and 1 week after treatment.Result:Five patients were lost to follow up (all five in the modified Semont group),and three patients failed to complete treatment (all three in the Epley group). The sequelae at the 3rd day and one week after modified Semont maneuver were 27 and 9,while 41 and 15 in Epley group. The efficacy rates at the 3rd day and one week after modified Semont maneuver were 91.7% and 98.3%,and 91.9% and 96.8% in Epley group retrospectively. The sequelae and short-term effective rate of patients in modified Semont group was no difference when compared with that in Epley group (P>0.05).Conclusion:Modified Semont maneuver represents a simple rapid and effective approach to the treatment of posterior canal benign paroxysmal positional vertigo.

[Current status of treatment of laryngeal contact granuloma].

Laryngeal contact granuloma, also known as vocal cord granuloma, is the non-neoplastic inflammatory granulation tissue occurring in the vocal cord around. The cause is unclear, morbidity and cure rate is low, the recurrence rate is high, leading to a poor clinical treatment. But some symptoms of the sound disorder, pharyngeal foreign body sensation seriously affect the quality of life of patients, increase the psychological burden of patients. With the increasing awareness of the disease, the deepening of the research, there have been different treatment methods. This paper reviews the previous literatures, and summarizes LCG treatment for the status.

[Analysis of the curative effect of intralesional steroid injection combined with acid-suppressive therapy for laryngeal contact granuloma].

OBJECTIVE: To study the efficacy of intralesional steroid injection combined with acid-suppressive therapy for laryngeal contact granuloma. METHODS: Eighty eight patients with laryngeal contact granuloma diagnosed from March 2010 to October 2014 were reviewed.Twenty-one patients took intralesional steroid injection combined with acid-suppressive therapy as experimental group (4 patients in initial treatment group, 17 patients in non-initial treatment group), other 67 patients only received acid-suppressive therapy as control group. The clinical data of these patients was analyzed. RESULTS: In experimental group, 15 patients were cured (71.43%) and the average cure time were (3.80±0.43) months, total injection time was 2-6 times and the median was 3 times. In control group, 50 patients were cured (74.63%) and the average cure time were (5.31±0.38) months. No any systemic and local complications were found in all the patients during the follow-up period of 6-42 months. The cure rate had no significant difference between the experimental group and control group, but the cure time in the former group was shorter than that in the latter group (Z=-2.093, P=0.036). In the experimental group, the initial treatment patients had no significant difference in cure rate and cure time compared with non initial patients (χ(2)=1.112, P=0.544; Z=0.000, P=1.000). CONCLUSION: Intralesional steroid injection combined with acid-suppressive therapy can be recommended as a safe and effective alternative therapy for laryngeal contact granuloma, especially the refractory granuloma.

[Diagnosis and management of pharyngeal dysphagia].

Objective:To evaluate the effects of different managements on the patients with pharyngeal dysphagia.Method:One hundred patients with pharyngeal dysphagia were recruited for this study.They were divided into three groups after evaluation of swallow function. Then the patients in each groups received treatments include oropharyngeal strengthening exercises, esophageal entrance balloon dilation and cricopharyngeal myotomy, respectively.After 3 months of treatment,the swallow function of the patients was evaluated again to determine the efficacy of the treatments.Result:Seventyfive patients received oropharyngeal strengthening exercises,21 patients received esophageal entrance balloon dilation and 4 patients received cricopharyngeal myotomy. After 3 months of treatment, the effective rates of three groups were 65.33%,76.19% and 75.00%, respectively, with a total efficacy of 68.00%. There was no significant difference of the treatment efficacy among three groups（χ²=0.983，P＞0.05）.Conclusion:Pharyngeal dysphagia can be improved after selected treatments according to the causes and severity of the disease.

Interfacial exchange coupling and magnetization reversal in perpendicular [Co/Ni]N/TbCo composite structures.

Interfacial exchange coupling and magnetization reversal characteristics in the perpendicular heterostructures consisting of an amorphous ferrimagnetic (FI) TbxCo(100-x) alloy layer exchange-coupled with a ferromagnetic (FM) [Co/Ni]N multilayer have been investigated. As compared with pure TbxCo(100-x) alloy, the magnetization compensation composition of the heterostructures shift to a higher Tb content, implying Co/Ni also serves to compensate the Tb moment in TbCo layer. The net magnetization switching field Hc⊥ and interlayer interfacial coupling field Hex, are not only sensitive to the magnetization and thickness of the switched TbxCo(100-x) or [Co/Ni]N layer, but also to the perpendicular magnetic anisotropy strength of the pinning layer. By tuning the layer structure we achieve simultaneously both large Hc⊥ = 1.31 T and Hex = 2.19 T. These results, in addition to the fundamental interest, are important to understanding of the interfacial coupling interaction in the FM/FI heterostructures, which could offer the guiding of potential applications in heat-assisted magnetic recording or all-optical switching recording technique.

Propofol and memory: a study using a process dissociation procedure and functional magnetic resonance imaging.

Thirty volunteers randomly received either mild or deep propofol sedation, to assess its effect on explicit and implicit memory. Blood oxygen level-dependent functional magnetic resonance during sedation examined brain activation by auditory word stimulus and a process dissociation procedure was performed 4 h after scanning. Explicit memory formation did not occur in either group. Implicit memories were formed during mild but not deep sedation (p = 0.04). Mild propofol sedation inhibited superior temporal gyrus activation (Z value 4.37, voxel 167). Deep propofol sedation inhibited superior temporal gyrus (Z value 4.25, voxel 351), middle temporal gyrus (Z value 4.39, voxel 351) and inferior parietal lobule (Z value 5.06, voxel 239) activation. Propofol only abolishes implicit memory during deep sedation. The superior temporal gyrus is associated with explicit memory processing, while the formation of both implicit and explicit memories is associated with superior and middle temporal gyri and inferior parietal lobule activation.

Effect of midazolam on memory: a study of process dissociation procedure and functional magnetic resonance imaging.

To assess the effects of midazolam on explicit and implicit memories, 12 volunteers were randomly divided into the two groups: one with an Observer's Assessment of Alertness/Sedation score of 3 (mild sedation) and one with a score of 1 (deep sedation). Blood oxygen-level-dependent functional magnetic resonance imaging was measured before and during an auditory stimulus, then with midazolam sedation, and then during a second auditory stimulus with continuous midazolam sedation. After 4 h, explicit and implicit memories were assessed. There was no evidence of explicit memory at the two levels of midazolam sedation. Implicit memory was retained at a mild level of midazolam sedation but absent at a deep level of midazolam sedation. At a mild level of midazolam sedation, activation of all brain areas by auditory stimulus (as measured by functional magnetic resonance imaging) was uninhibited. However, a deep level of midazolam sedation depressed activation of the superior temporal gyrus by auditory stimulus. We conclude that midazolam does not abolish implicit memory at a mild sedation level, but can abolish both explicit and implicit memories at a deep sedation level. The superior temporal gyrus may be one of the target areas.