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This study aimed to investigate the effect of treadmill exercise on neuropathic pain and to determine whether mitophagy of the anterior cingulate cortex (ACC) contributes to exercise-mediated amelioration of neuropathic pain. Chronic constriction injury of the sciatic nerve (CCI) was used to establish a neuropathic pain model in Sprague-Dawley (SD) rats. Von-Frey filaments were used to assess the mechanical paw withdrawal threshold (PWT), and a thermal radiation meter was used to assess the thermal paw withdrawal latency (PWL) in rats. qPCR was used to evaluate the mRNA levels of Pink1, Parkin, Fundc1, and Bnip3. Western blot was used to evaluate the protein levels of PINK1 and PARKIN. To determine the impact of the mitophagy inducer carbonyl cyanide m-chlorophenylhydrazone (CCCP) on pain behaviors in CCI rats, 24 SD rats were randomly divided into CCI drug control group (CCI+Veh group), CCI+CCCP low-dose group (CCI+CCCP0.25), CCI+CCCP medium-dose group (CCI+CCCP2.5), and CCI+CCCP high-dose group (CCI+CCCP5). Pain behaviors were assessed on 0, 1, 3, 5, and 7 days after modeling. To explore whether exercise regulates pain through mitophagy, 24 SD rats were divided into sham, CCI, and CCI+Exercise (CCI+Exe) groups. The rats in the CCI+Exe group underwent 4-week low-moderate treadmill training one week after modeling. The mechanical pain and thermal pain behaviors of the rats in each group were assessed on 0, 7, 14, 21, and 35 days after modeling. Western blot was used to detect the levels of the mitophagy-related proteins PINK1, PARKIN, LC3 II/LC3 I, and P62 in ACC tissues. Transmission electron microscopy was used to observe the ultrastructure of mitochondrial morphology in the ACC. The results showed that: (1) Compared with the sham group, the pain thresholds of the ipsilateral side of the CCI group decreased significantly (P < 0.001). Meanwhile, the mRNA and protein levels of Pink1 were significantly higher, and those of Parkin were lower in the CCI group (P < 0.05). (2) Compared with the CCI+Veh group, each CCCP-dose group showed higher mechanical and thermal pain thresholds, and the levels of PINK1 and LC3 II/LC3 I were elevated significantly (P < 0.05, P < 0.01). (3) The pain thresholds of the CCI+Exe group increased significantly compared with those of the CCI group after treadmill intervention (P < 0.001, P < 0.01). Compared with the CCI group, the protein levels of PINK1 and P62 were decreased (P < 0.001, P < 0.01), and the protein levels of PARKIN and LC3 II/LC3 I were increased in the CCI+Exe group (P < 0.01, P < 0.05). Rod-shaped mitochondria were observed in the ACC of CCI+Exe group, and there were little mitochondrial fragmentation, swelling, or vacuoles. The results suggest that the mitochondrial PINK1/PARKIN autophagy pathway is blocked in the ACC of neuropathic pain model rats. Treadmill exercise could restore mitochondrial homeostasis and relieve neuropathic pain via the PINK1/PARKIN pathway.
Subject(s)
Rats , Animals , Mitophagy/physiology , Rats, Sprague-Dawley , Carbonyl Cyanide m-Chlorophenyl Hydrazone/pharmacology , Gyrus Cinguli , Neuralgia , Ubiquitin-Protein Ligases/metabolism , Protein Kinases , Membrane Proteins/metabolism , Mitochondrial Proteins/metabolismABSTRACT
Objective:To explore the association rules of personality traits and health-promoting lifestyle in patients with primary angle closure glaucoma, so as to provide advice for the synthetical treatment.Methods:From July to November 2021, a total of 117 primary angle closure glaucoma patients(acute patients n=89, chronic patients n=28) in ophthalmology department of five hospitals in Nanjing were investigated with type A behavior pattern scale, health-promoting lifestyle scale Ⅱ and general information questionnaire.Based on Weka 3.8.5, algorithm of Apriori was used to mine its association relationship. Results:(1) The total scores of type A behavior pattern scale for patients with acute and chronic types of primary angle closure glaucoma were (32.48±6.43) and (27.54±6.49) respectively.The total scores of health-promoting lifestyle scale Ⅱ were (101.69±11.83) and (97.79±7.78) respectively.(2) There were positive associations among patients with acute primary angle closure glaucoma, type A/A-personality (including impatience and hostility) and health-promoting lifestyle (including stress management disorder, interpersonal relationship management disorder, well sense of health responsibility and adequate dietary nutrition intake)(all support>0.1, confidence >0.6, lift >1.0). And patients with chronic primary angle closure glaucoma were associated with B/B-personality (including patience and mild), health-promoting lifestyle (including stress management disorder, interpersonal relationship management disorder, well sense of health responsibility and adequate dietary nutrition intake)(all support>0.1, confidence >0.6, lift >1.0).Conclusion:Primary angle closure glaucoma is strongly related with personality traits and health-promoting lifestyle.Its synthetical treatment plan should take both physical and mental measures, and classified health management for patients with different disease types.
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Objective:To analyze the clinical features of absolute incidental prostate cancer (AIPCa) and pseudo-incidental prostate cancer (PIPCa).Methods:Between January 2013 and October 2019, 52 male patients who were diagnosed as incidental prostate cancer (IPCa) with their postoperative pathological examination in our center were included. None of them had any evidence of prostate cancer before. Their median age was 77.5(ranging 55-93) years old. 48 cases underwent PSA examination. The median tPSA was 6.7 (ranging 1-46) ng/ml, except 1 case tPAS>100ng/ml. The median IPSS score was 27.7(ranging 12-35). 48 cases accepted prostatic ultrasound examination. The median prostate volume was 38.6 (ranging 2.3-130.2)ml. Among them, 13 patients underwent transurethral resection of the prostate (TURP), 24 patients underwent transurethral enucleation and resection of the prostate(TUERP), 15 patients underwent radical cystoprostatectomy(RCP). According to the patients′ preoperative examination, AIPCa and PIPCa were redefined. Clinical characteristics, postoperative pathology and follow-up of the two groups were compared and analyzed.Results:There was no significant difference in age, IPSS score, tPSA, T stage and Gleason score between the operation methods groups except for prostate volume and resection ration of prostate ( P>0.05). Among those group, the sequence of prostatic resection ration was RCP>TUERP>TURP( P<0.001). There was no significant difference in age, IPSS score, prostate volume, proportion of T 1b and Gleason score between AIPCa and PIPCa groups. However, the analysis showed higher tPSA(14.68 ng/ml vs. 3.14 ng/ml) in PIPCa ( P<0.001). With a mean follow-up of 33 months(ranging 6-78 months), 23.8%(5/21) patients in AIPCa group and 45.2%(14/31) patients in PIPCa group were found to have increased PSA and underwent radical prostatectomy or androgen deprivation therapy in the follow-up. There was no metastasis or death in AIPCa group, while 2 patients had bone metastasis and 1 patient died of prostate cancer in PIPCa group. Conclusions:According to the indication of prostate biopsy, PIPCa and AIPCa were defined. PIPCa has a relatively poor prognosis, some of them may be missed diagnosis and clinical underestimation. In order to avoid missed diagnosis before operation, we should strictly define IPCa and follow the indications of biopsy, emphasize the reference value of PSA in tumor screening and biopsy, and careful handle the negative results of biopsy when PSA is abnormal.
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Objective:To investigate the effect of sodium arsenite (NaAsO 2) on transcriptional activity of nuclear factor E2-related factor 2 (Nrf2) signaling pathway in mouse lymph node vascular endothelial cell line (SVEC4-10). Methods:In vitro cell culture method was used to treat SVEC4-10 cells for 24 h with different doses of NaAsO 2 [0 (control), 2, 5, 10, 20, 50, 100, 150 μmol/L], and the cell viability was detected by tetrazole compound (MTS) method. The time-response relationship was studied with SVEC4-10 cells treated with 5 μmol/L NaAsO 2 for 0 (control), 2, 6 and 12 h; the dose-response relationship was studied with SVEC4-10 cells treated with 0 (control), 2, 5 and 10 μmol/L NaAsO 2 for 6 h; real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the mRNA expression of Nrf2 and its downstream genes glutamate-cysteine ligase catalytic subunit (Gclc), glutamate-cysteine ligase modifier subunit (Gclm), NAD(P)H dehydrogenase quinone 1 (Nqo1) and metallothionein 1 (Mt1). Establishment of Nrf2 gene stably silenced (Nrf2-KD) cells using SVEC4-10 cells, the interference control (scramble, SCR) cells and Nrf2-KD cells were treated with 0(control), 10 and 20 μmol/L NaAsO 2 for 16 h, and apoptosis was detected by flow cytometry. Results:MTS test results showed that the cell viability of the control, 2, 5, 10, 20, 50, 100, 150 μmol/L NaAsO 2 treatment groups was (100.00 ± 19.53)%, (98.18 ± 9.85)%, (96.09 ± 30.04)%, (90.64 ± 8.74)%, (59.75 ± 12.09)%, (35.43 ± 8.58)%, (26.35 ± 5.89)% and (17.54 ± 4.48)%, respectivily. There was statistically significant difference in cell viability between different dose groups ( F = 18.30, P < 0.05); and the cell viability of the 20, 50, 100, 150 μmol/L NaAsO 2 treatment groups was significantly lower than that of the control group ( P < 0.05). The time-response relationship results showed that there were statistically significant differences in Nrf2, Gclc, Gclm, Nqo1 and Mt1 mRNA level between control, 2, 6 and 12 h treatment groups ( F = 56.69, 85.28, 90.82, 80.46, 758.60, P < 0.05); with extension of arsenic exposure time, the mRNA level of Nrf2, Gclc, Gclm and Mt1 first increased and then decreased, the mRNA level of Nqo1 increased continually; among them, the mRNA level of Nrf2 peaked at 2 h, the mRNA levels of Gclc, Gclm and Mt1 peaked at 6 h, and the mRNA level of Nqo1 peaked at 12 h. The dose-response relationship results showed that there were statistically significant differences in Nrf2, Gclc, Gclm, Nqo1 and Mt1 mRNA levels between control, 2, 5 and 10 μmol/L NaAsO 2 treatment groups ( F = 68.39, 72.26, 30.41, 397.00, 28.88, P < 0.05); with increasing of arsenic exposure dose, the mRNA levels of Nrf2, Gclc, Gclm, Nqo1 and Mt1 increased. The mRNA level of Nrf2 peaked at a dose of 5 μmol/L, and the mRNA levels of Gclc, Gclm, Nqo1 and Mt1 peaked at a dose of 10 μmol/L. Apoptosis test results showed that there were statistically significant differences in the apoptosis rates of SCR and Nrf2-KD cells between control, 10 and 20 μmol/L NaAsO 2 treatment groups ( F = 8.18, 9.66, P < 0.05); compared with the control group, the apoptosis rates of SCR and Nrf2-KD cells in the 20 μmol/L NaAsO 2 treatment group increased ( P < 0.05); and the apoptosis rate of Nrf2-KD cells in the 20 μmol/L NaAsO 2 treatment group was higher than that of SCR cells in the same dose group ( P < 0.05). Conclusions:NaAsO 2 exposure has caused the activation of Nrf2 signaling pathway in mouse lymph node vascular endothelial cell line SVEC4-10 cells, activated the adaptive antioxidant response, and altered transcriptional activity; while silence of Nrf2 has made SVEC4-10 cells more sensitive to NaAsO 2 toxicity.
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Objective To investigate the protective effect of intravitreal injection of nerve growth factor (NGF) on apoptosis of retinal ganglion cells (RGCs) in early diabetic rats and its possible mechanism.Methods SD rats were divided into normal control group,diabetic model group,phosphate buffer solution (PBS) group and NGF group according to random number table method,with each group contain 6 rats.The rats in diabetic model group,PBS group and NGF group were injected intraperitoneally with streptozotocin (STZ) to establish diabetic rat model.After 4 weeks of modeling,2 μl PBS and 2 μl NGF (0.5 μg/μl) were injected into the vitreous cavity in PBS group and NGF group,respectively.The right eyes served as the experimental eyes,inject once a week for 4 weeks.After 4 weeks of injection,the retina microangiopathy of each group was observed by Phoenix Micron Ⅳ small animal retinal imaging system.After high dose anesthesia,the eyeball of one rat from each group was taken to prepare ultrathin sections and observed by transmission electron microscopy.Five rat eyeballs were taken to prepare retinal paraffin sections from each groups.The RGC apoptotic index was observed by TUNEL method.The expressions of RGC bcl-2 protein and bax protein were analyzed by immunohistochemistry.This study was approved by the Experimental Animal Ethics Committee of Central South University,and the experimental procedures were in accordance with the National Institutes of Health(NIH) guidelines for the Care and Use of Laboratory,and follow the 3R principle.Results The weight of the diabetic model rats was significantly lower than that of the normal group,and the intake of water and food were significantly increased,the urine volume was also in creased.The body weight and blood glucose level of the rats were significantly different among different groups after 4 weeks of modeling (F=202.352,148.444,both at P＜0.001).Fundus photography of early diabetic rats showed no obvious diabetic retinal microangiopathy.The numbers of RGC in the diabetic model group and PBS group were significantly lower than that in the normal control group under the transmission electron microscope.The membrane shrinkage,cytoplasmic condensation,organelle edema,chromatin peripheral collection were obvious and the cell apoptosis number was increased.The RGC lesions in the NGF group were lighter than those in the diabetic model group and PBS group.The apoptotic indexs in the normal control group,diabetic model group,PBS group and NGF group were (3.88±1.28)％,(92.56±1.58)％,(92.64±2.30)％ and (59.34±3.89)％,respectively,the overall difference of apoptotic index between the four groups was statistically significant (F=854.554,P＜0.001);the RGC apoptotic index of the diabetic model group was significantly higher than that of the normal control group.The RGC apoptotic index of the NGF group was significantly lower than that of the diabetic model group and PBS group (both at P＜0.05).The expression levels of bcl-2 protein in normal control group,diabetic model group,PBS group and NGF group were 38.11 ± 1.01,22.38±3.90,23.04±3.14 and 84.69±1.45,respectively,with a significant difference among the groups (F =366.206,P＜0.001).The expression levels of bax protein in normal control group,diabetes model group,PBS group and NGF group were 4.22± 1.89,56.59±6.67,56.30±8.51 and 26.19±2.44,respectively,with a significant difference among the groups (F=61.435,P＜0.001).The expression of bcl-2 protein in the diabetic model group was significantly lower than that in the normal control group (P＜0.05),and the expression of bax protein was significantly increased (P＜0.05).The expression of bcl-2 protein in NGF group was significantly higher and the expression of bax protein was significantly lower than those in the PBS group and diabetic model group (all at P＜0.05).Conclusions The ultrastructural damage of RGCs has occurred in early diabetic rats without diabetic retinal microangiopathy.Intravitreal injection of NGF may produce retinal neuroprotective effects by inhibiting apoptosis of RGCs in diabetic rats.
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Objective@#To investigate the effect of triptolide, a specific inhibitor of heat shock protein 70 (HSP70), on apatinib resistance in gastric cancer cells line MKN45.@*Methods@#The apatinib-resistant cells (MKN45/AR) and MKN45 parental cells were treated with apatinib, triptolide and apatinib combined with triptolide, respectively. CCK-8 assay was performed to determine the half maximal inhibitory concentration (IC50) of MKN45/AR and MKN45 cells in the presence of different treatment. The mRNA expression of heat shock protein gene (HSPA1A and HSPA1B) was detected by RT-PCR, while the protein expression of heat shock protein 70 was analyzed using Western blot in MKN45/AR and MKN45 cells.@*Results@#The IC50 values of apatinib-sensitive and apatinib-resistant MKN45 cells were 10.411 μmol/L and 70.527 μmol/L, respectively, showing a significant difference (P<0.05). The mRNA expression of HSPA1A and HSPA1B in MKN45/AR cells was significantly higher than that in MKN45 cells (P<0.001). The protein expression of heat shock protein 70 was significantly decreased after 0.25 μmol/L triptolide treatment in MKN45/AR cells (P<0.01). When heat shock protein 70 was inhibited by triptolide, the IC50 value of apatinib in MKN45/AR cells was reduced to 11.679 μmol/L, which was significantly lower than cells treated with apatinib alone (P<0.05).@*Conclusions@#The apatinib-resistant MKN45 cells have high levels of heat shock protein 70. Low doses of triptolide can significantly inhibit heat shock protein 70, leading to reverse the resistance phenotype of MKN45/AR cells. Therefore, inhibition of heat shock protein 70 provides a new therapy strategy for patients with apatinib resistance.
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Objective To explore the association rules of behavior problems and parenting styles in asthmatic children. Methods The database was established with the data of behavior problems and paren-ting styles of 202 allergic asthma children.Based on the algorithm of Apriori,the relativity between these be-havior problems and parenting styles was mined.314 healthy children of the same age were selected as the control group. Results (1)The differences in behavior problems between asthmatic group and control group were significant,which includes attention(7.63±3.44 vs 4.03±3.01),sociality(4.10±2.14 vs 2.86± 1.67),aggression(11.74±5.10 vs 6.63±4.26)and flinch(3.03±1.58 vs 2.06±1.38)(all P<0.01),while the difference in offence between two groups was not significant(P>0.05).The differences in parenting styles be-tween asthmatic group and control group were significant,including spoiledness(15.26 ± 4.16 vs 10.88 ± 3.29),indulgence(22.47±4.68 vs 16.59±4.12),discrepancy(13.54±3.50 vs 11.92±3.35)and democracy (35.08±5.25 vs 39.76±4.80)(all P<0.01),while the difference in autocracy was not significant between two groups(P>0.05).(2)There were multi-positive associations among these behavior problems of asthmatic children as followed.There were positive associations between the cognitive impairment(such as attention) and social impairment(including flinch,aggression and sociality),(each support>10%,confidence>0.60,lift>1.20,the value of correlation coefficient between(1,+∞)).(3)There were multiple associations between behavior problems and parenting styles of asthmatic children as followed.There were negative associations be-tween democracy and behavior problems(the value of correlation coefficient between[0,1)),and there were positive associations among indulgence、spoiledness,discrepancy and these behavior problems(the value of correlation coefficient between(1,+∞)).Conclusion There are positive associations among these behavior problems of asthmatic children,and there are multi-associations between behavior problems and parenting styles.
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Objective To explore the effect of parenting style and behavior on children with allergic asthma.Methods From July to December in 2016,a total of 60 children with allergic asthma,who were from Jiangsu Province Hospital of Traditional Chinese Medicine and Jiangsu Research Institute of Traditional Chinese Medicine,were selected as the asthma group.Another 60 normal children from kindergarten and primary school were selected as the healthy group.Achenbach child behavior checklist (CBCL) and parenting style questionnaire were used to evaluate the behavior and parenting style of the two groups.The Fractional exhaled nitric oxide (FeNO) of the two groups were also collected.Results (1) The scores of dotage,autocracy,inconsistence of parenting style in asthma group ((14.95±4.95),(22.23±4.69),(13.43±3.80))were higher than those in healthy group((11.68±3.69),(16.47±4.16),(11.07±3.32))and the differences were significant (all P＜0.05).The scores of democracy,indulgence in asthma group ((38.92 ± 5.37),(18.12± 4.54)) were lower than those in healthy group((41.35±3.62),(19.88±3.41)),and the differences were statistically significant(all P＜0.05).(2) The scores of attention,socialization,aggression and withdrawal of child behavior in asthma group ((7.35±3.55),(3.85±2.17),(11.57±5.30),(3.40±2.65))were higher than those in healthy group((3.38±2.81),(2.52± 1.52),(6.98±4.77),(2.02± 1.63)),the differences were statistically significant(all P＜0.01).There was no significant difference between the two groups in the dimension of discipline(P＞0.05).(3) The FeNO index of asthma group((32.27± 14.27) × 10-9 mol/L) was higher than that of healthy group ((11.10± 1.88) × 10-9 mol/L) and the differences were statistically significant (P＜0.01).(4)Logistic regression analysis showed that the attention(B=0.38,OR=1.47,95％ CI =1.27-1.70,P＜0.01),dotage(B=0.19,OR=1.20,95％ CI =1.06-1.37,P＜0.01) and autocracy(B=0.24,OR=1.27,95％ CI =1.14-1.42,P＜0.01)had significant effects on allergic asthma.Conclusion Attention,dotage and autocracy are the risk factors to influence the control level of allergic asthma.This should arouse the attention of clinicians and parents.
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<p><b>OBJECTIVES</b>To study the correlation between MRI apparent diffusion coefficient (ADC) and expression of Ki-67 in gastric cancers, and to investigate the application of ADC value in diagnosing the malignance of gastric cancer.</p><p><b>METHODS</b>A retrospective cohort analysis was performed on 87 gastric cancer patients who received MRI examination and radical resection at Zhejiang Provincial Hospital of Traditional Chinese Medicine from November 2014 to August 2015. All the postoperative resected samples were confirmed as gastric cancer. Preoperative MRI examination was performed by using Siemens 3.0-T Verio MRI with following parameters: section thickness 3 mm, gap 1 mm, matrix 182×320, field of view 40 cm. Plain scan was followed by T1-weighted fat suppression technique VIBE 3D(TR3.92/TE1.39,90degree) scans at arterial phase (the 30th second), portal venous phase (the 60th second), lag period (the 90th second), axial planes and coronal planes (the 180th second), and sagittal planes (the 210th second), respectively. ADC value of tumor was measured at b-factor of 800 s/mmand ADC map was generated from DWI data on the work station. The expression of Ki-67 in cancer tissue was detected by routine immunohistochemical (SP) staining after surgery. Correlation between ADC value and the expression of Ki-67 in gastric cancer was analyzed.</p><p><b>RESULTS</b>Irregular thickening of the gastric wall and inhomogeneous enhancement of the tumor after injection of the contrast material appeared in gastric cancer. Gastric cancer tissue presented hyperintensity and normal gastric wall presented isointensity in DWI image (b=800 s/mm). Compared with normal gastric tissue, mean ADC value of gastric cancer tissue was significant lower [(1.114±0.265)×10mm/s vs. (2.032±0.202)×10mm/s, t=26.209, P=0.000]. The ADC values of high-middle differentiation group, middle-low differentiation group, low differentiation group and signet ring cell carcinoma/mucinous adenocarcinoma group were (1.347±0.234)×10mm/s, (1.179±0.257)×10mm/s, (0.996±0.185)×10mm/s and (1.082±0.230)×10mm/s, respectively. The difference of mean ADC value among different tumor stages was significant(F=8.498, P=0.000). Along with the Ki-67 expression up-regulated, the ADC value decreased in cancer tissue. The Ki-67 expressions in cancer tissue was negatively correlated with cancer ADC values (r=-0.570, P=0.000). Furthermore, negative correlations of Ki-67 expressions with ADC values of high-middle differentiation group (r=-0.627, P=0.016), low differentiation group (r=-0.787, P=0.000) and signet ring cell carcinoma/mucinous adenocarcinoma group (r=-0.792, P=0.000) were observed respectively, while Ki-67 expression was not correlated with ADC value of middle-low differentiation group.</p><p><b>CONCLUSION</b>The ADC value of gastric cancer can reflect the level of tumor differentiation, and is negatively correlated with Ki-67 expression in cancer tissues.</p>