Role and mechanism of interleukin-8 in bone regeneration / 中国组织工程研究

BACKGROUND:Interleukin-8 is an important cytokine that has been found to play an important role in bone regeneration through multiple pathways. OBJECTIVE:To comprehensively review the action mechanism of interleukin-8 effects on bone regeneration to provide ideas for the following studies on interleukin-8. METHODS:By searching the China National Knowledge Infrastructure database for articles published from January 1999 to February 2023 and PubMed database for articles published from January 1985 to February 2023 reporting the role of interleukin-8 in bone-associated cells and vascularisation.Chinese and English search terms were"interleukin-8,bone repair,bone metabolism,mesenchymal stem cells,osteoblasts,osteoclasts,vascularization".The initial review yielded 508 articles in English and Chinese,and a total of 51 articles were included for review and analysis according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION:According to the existing research,interleukin-8 can promote bone cell regeneration and assist bone healing through multiple pathways,which is mainly divided into three aspects:(1)Promote the proliferation and differentiation of bone cells such as mesenchymal stem cells and osteoblasts,and promote the development of cells in the direction of promoting bone healing;(2)interleukin-8 can promote angiogenesis and provide sufficient nutrition and oxygen for bone tissue,thus further improving the quality and stability of bone healing.(3)The appearance of interleukin-8 facilitates the expression of hypoxia-inducible factor-1α,vascular endothelial growth factor,and matrix metalloproteinase,which can create a microenvironment conducive to bone regeneration,thus promoting the regeneration and repair of bone tissue.In summary,interleukin-8 plays an important role in bone healing by promoting osteoblast proliferation and differentiation,facilitating angiogenesis and improving the mechanical properties of bone regeneration,as well as influencing bone metabolism through osteoclasts,mesenchymal stem cells,and other action sites.

Impairment of Autophagic Flux After Hypobaric Hypoxia Potentiates Oxidative Stress and Cognitive Function Disturbances in Mice / 神经科学通报·英文版

Acute hypobaric hypoxic brain damage is a potentially fatal high-altitude sickness. Autophagy plays a critical role in ischemic brain injury, but its role in hypobaric hypoxia (HH) remains unknown. Here we used an HH chamber to demonstrate that acute HH exposure impairs autophagic activity in both the early and late stages of the mouse brain, and is partially responsible for HH-induced oxidative stress, neuronal loss, and brain damage. The autophagic agonist rapamycin only promotes the initiation of autophagy. By proteome analysis, a screen showed that protein dynamin2 (DNM2) potentially regulates autophagic flux. Overexpression of DNM2 significantly increased the formation of autolysosomes, thus maintaining autophagic flux in combination with rapamycin. Furthermore, the enhancement of autophagic activity attenuated oxidative stress and neurological deficits after HH exposure. These results contribute to evidence supporting the conclusion that DNM2-mediated autophagic flux represents a new therapeutic target in HH-induced brain damage.

Protective effect of Dictyophora polysaccharide on neurotoxicity induced by sodium arsenite in rats / 中华地方病学杂志

Objective:To investigate the protective effect of Dictyophora polysaccharide on neurotoxicity induced by sodium arsenite in rats.Methods:Sixty SD rats (half males and half females) were selected and fed adaptively for one week. The rats were divided into a normal group ( n = 20, ordinary feed) and a modeling group [ n = 40, arsenic-containing feed (50 mg/kg sodium arsenite)] according to their body weight (80 - 100 g) by random number table method. After 12 weeks, the arsenic content in brain and blood of the rats ( n = 10) was measured to identify the arsenism model. After successful modeling, the rats in the modeling group were divided into Dictyophora polysaccharide group (arsenic-containing feed + 20 ml·kg -1·bw Dictyophora polysaccharide solution by gavage), and model group (arsenic-containing feed + equal volume distilled water by gavage), while the rats in the normal group (ordinary feed + equal volume distilled water by gavage) were retained, with 10 rats in each group for 4 weeks of intervention. Morris water maze test was used to assess the spatial learning and memory ability of the rats. Nissl staining was used to observe the pathological changes of the brain tissue, and the oxidative stress factors [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA)], and inflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin -1β (IL-1β)] in the brain tissue of each group were measured. Results:Brain arsenic content of rats in the modeling group [(92.02 ± 13.37) μg/g] and blood arsenic content [(51.37 ± 19.33) μg/L] were higher than those of the normal group [(7.42 ± 3.21) μg/g and (2.74 ± 1.29) μg/L, t = - 6.91, - 6.06, P < 0.001]. The rat model of arsenic poisoning was successfully established. Compared with the normal group, the escape latency on the 1st, 3rd and 4th day and the first arrival time of rats in the model group were prolonged, the number of platform crossings was reduced, and the proportion of target quadrant residence time was decreased ( P < 0.05). Compared with the model group, the escape latency on the 4th day and the first arrival time of rats in the Dictyophora polysaccharide group were shortened, and the proportion of target quadrant residence time was prolonged ( P < 0.05). The results of Nissl staining showed that compared with the normal group, the number of Nissl bodies was decreased, the intercellular space increased, and the arrangement was disorderly in the model group; compared with the model group, the number of Nissl bodies was increased, most of the neurons were structurally intact. Compared with the normal group, the levels of SOD and GSH-Px in the brain tissue of rats in the model group were lower, and the levels of MDA, TNF-α and IL-1β were significantly higher ( P < 0.05). Compared with the model group, the levels of SOD and GSH-Px in the brain tissue of rats in the Dictyophora polysaccharide group were higher, while the levels of MDA, TNF-α and IL-1β were lower ( P < 0.05). In addition, the levels of SOD, GSH-Px, MDA and IL-1β in the Dictyophora polysaccharide group were not significantly different from those in the normal group ( P > 0.05). Conclusion:Diactyophora polysaccharide probably reduces the neurotoxicity damage caused by sodium arsenite in rats through antioxidant and antiinflammatory effects.

Expression of 5-methylcytosine and 5-hydroxymethylcytosine in bone tissue of rats with different types of skeletal fluorosis / 中华地方病学杂志

Objective:To learn about the levels of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in bone tissue of rats with different types of skeletal fluorosis and analyze their correlation.Methods:Thirty 4-week-old SPF grade healthy SD rats were selected. After adaptive feeding for 1 week, the rats were divided into control group (4 ml·kg -1·bw deionized water + standard maintenance diet), osteosclerosis group [20 mg·kg -1·bw sodium fluoride (NaF) + standard maintenance diet], and osteoporosis/osteomalacia group (20 mg·kg -1·bw NaF + low-calcium and low-protein partial diet) according to their body weight (100 - 120 g) by random number table method, with 10 rats in each group, half male and half female; gavaged 6 days each week and the experimental period was 5 months. At the end of the experiment, samples of rat heart blood and lower limb femur were collected. The contents of serum methyl donor S-adenosylmethionine (SAM) and its metabolite S-adenosylhomocysteine (SAH) in serum, and the levels of 5-mC and 5-hmC in bone tissue were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to determine the expression of DNA methyltransferase (DNMTs) and DNA hydroxymethylase (TETs) in bone tissue of rats. The correlation between serum SAM content, SAM/SAH ratio and bone tissue 5-mC level, and between the bone tissue 5-mC level and 5-hmC level was analyzed. Results:Serum SAM [11.03 (7.06, 18.63), 3.96 (2.32, 9.09), 3.91 (2.35, 4.46) nmol/L], SAH content [(4.69 ± 0.55), (5.41 ± 1.13), (13.90 ± 1.09) ng/L], SAM/SAH ratio [2.58 (1.54, 4.12), 0.62 (0.52, 1.69), 0.14 (0.13, 0.15)] and bone tissue 5-mC [103.39 (97.37, 109.35), 52.50 (50.19, 68.13), 55.03 (49.97, 59.57) ng/L], 5-hmC levels [(32.61 ± 8.84), (56.96 ± 8.48), (20.34 ± 6.22) ng/L] in the control group, osteosclerosis group and osteoporosis/osteomalacia group were compared, and the differences were statistically significant beween three groups ( H/ F = 12.81, 284.24, 21.85, 19.37, 55.23, P < 0.01). Compared with the control group, the content of SAM, the ratio of SAM/SAH, the level of 5-mC in the osteosclerosis group and osteoporosis/osteomalacia group, and the level of 5-hmC in the osteoporosis/osteomalacia group were lower ( P < 0.05), while the content of SAH in the osteoporosis/osteomalacia group and the level of 5-hmC in the osteosclerosis group were higher ( P < 0.05). Compared with the osteosclerosis group, the content of SAH in the osteoporosis/osteomalacia group was higher, while the ratio of SAM/SAH and the level of 5-hmC were lower ( P < 0.05). Western blot showed that there were statistically significant differences in the expression levels of DNMT1, DNMT3A, DNMT3B, TET1 and TET2 protein in bone tissue of rats in the control group, osteosclerosis group, and osteoporosis/osteomalacia group ( F = 285.45, 345.58, 239.83, 311.52, 318.24, P < 0.001). Among them, the expression levels of DNMT1, DNMT3A and DNMT3B protein in the osteosclerosis group and osteoporosis/osteomalacia group were lower than those in the control group, and the expression levels of DNMT1, DNMT3A and DNMT3B protein in the osteosclerosis/osteomalacia group were lower than those in the osteosclerosis group ( P < 0.05); the expression levels of TET1 and TET2 protein in osteosclerosis group were higher than those in the control group and osteoporosis/osteomalacia group, and the expression levels of TET1 and TET2 protein in the osteoporosis/osteomalacia group were lower than those in the control group ( P < 0.05). The results of Spearman rank correlation analysis showed that the content of SAM and the ratio of SAM/SAH in the control group, osteosclerosis group and osteoporosis/osteomalacia group were positively correlated with the level of 5-mC in bone tissue ( rs = 0.89, 0.92, 0.81, 0.73, 0.87, 0.73, P < 0.05). The levels of 5-mC and 5-hmC in bone tissue of rats in each group were negatively correlated ( rs = - 0.69, - 0.68, - 0.72, P < 0.05). Conclusions:The level of 5-mC in bone tissue of osteosclerotic fluorosis rats is low, and the level of 5-hmC is high, while those of osteoporosis/osteomalacia fluorosis rats are lower. The difference of 5-mC level in bone tissue of rats with different types of skeletal fluorosis is not significant, which may be related to the difference of 5-hmC level in bone tissue.

Analysis on risk factors for prognosis of traumatic brain injury in adults and establishment of the prediction model / 中华创伤杂志

Objective:To analyze risk factors for prognosis of adult patients with traumatic brain injury (TBI), construct the prognostic model of TBI and evaluate its predictive value.Methods:A case-control study was used to analyze the clinical data of 522 patients with TBI admitted to Xijing Hospital of Air Force Medical University from March 2011 to September 2019, including 438 males and 84 females; aged 18-75 years [(44.9±15.0)years]. According to the Glasgow outcome score (GOS) at discharge, the patients were divided into good prognosis group (GOS 4-5 points, n=165) and poor prognosis group (GOS 1-3 points, n=357). The two groups were compared with regards to qualitative data such as sex, underlying diseases, causes of injury, multiple injuries, open injuries, intracranial foreign bodies, cerebral herniation, consciousness status on admission and at discharge, surgery, lung infection on admission, tracheostomy, ventilator-assisted ventilation, hospital-acquired pneumonia/pathogenic bacteria and intracranial infection, and quantitative data such as Glasgow coma score (GCS) on admission and at discharge, age, measurements on admission [systolic blood pressure, diastolic blood pressure, mean arterial pressure, temperature, heart rate, creatinine, urea nitrogen, blood sodium, blood potassium, blood glucose, prothrombin time (PT), activated partial thromboplastin time (APTT), platelets, international normalized ratio (INR), pupil size of both eyes] and length of hospital stay. Univariate analysis and Lasso regression analysis were used to screen the risk factors affecting the prognosis of TBI patients, and the selected influencing factors were included in multivariate Logistic regression analysis to identify independent risk factors and construct regression equations. R was used to draw a visual nomogram based on regression equation for predicting the prognosis of TBI patients. The prognostic predictive value of the nomogram was evaluated by using the receiver operating characteristic (ROC) curve, and the area under the curve (AUC), Youden index, sensitivity, specificity and consistency index (C index) were calculated. Results:Univariate analysis showed that there were significant differences between the two groups in underlying diseases, open injuries, cerebral herniation, consciousness status on admission and at discharge, lung infection on admission, tracheostomy, ventilator-assisted ventilation, hospital-acquired pneumonia/pathogenic bacteria, GCS on admission and at discharge, age, and measurements on admission (systolic blood pressure, mean arterial pressure, body temperature, heart rate, creatinine, urea nitrogen, blood potassium, blood glucose, PT, INR, pupil size of right eye) (all P<0.05 or 0.01). There were no significant differences between the two groups in gender, causes of injury, multiple injuries, intracranial foreign bodies, surgery, intracranial infection, measurements on admission (diastolic blood pressure, blood sodium, APTT, platelets, pupil size of left eye) and length of hospital stay (all P>0.05). After screening by Lasso regression model, the results of multivariate Logistic regression analysis showed that GCS on admission ( OR=0.67, 95% CI 0.62, 0.73, P<0.01), age ( OR=1.03, 95% CI 1.01, 1.04, P<0.01), blood glucose on admission ( OR=1.17, 95% CI 1.06, 1.30, P<0.01) and INR on admission ( OR=17.08, 95% CI 2.12, 137.89, P<0.01) could be used as the main risk factors to construct the prediction model, and the regression equation was constructed: Logit [ P/(1- P)]=-0.398× "GCS on admission"+0.024× "age"+0.158×"blood glucose on admission"+2.838×"INR on admission"-1.693. The AUC for the prognosis prediction in adult patients with TBI using R based on a visual nomogram model was 0.87 (95% CI 0.83, 0.89, P<0.01). The Youden index for the predicted probability was 0.60 (sensitivity of 85.2% and specificity of 75.2%), with the C index of 0.87. Conclusion:Age, GCS on admission, blood glucose on admission and INR on admission are the main risk factors affecting the prognosis of TBI in adults, and the nomogram drawn by these parameters can better predict their clinical outcome.

Discussion on the staged treatment of chronic heart failure based on the theory of "deficiency, blood stasis, water and toxin" / 国际中医中药杂志

Deficiency, stasis, water and toxin are of great significance in the pathogenesis and pathologic evolution of chronic heart failure (CHF). Based on "deficiency, blood stasis, water and toxin", the pathogenesis and treatment of CHF were discussed in this article. It was found that in the pathogenesis, deficiency--deficiency of heart qi and deficiency of heart yang were the origin of the disease, and blood stasis, water and toxin were the markers of the disease. Among them, blood stasis was the central pathological link, and also an important mechanism that could aggravate the disease and cause a vicious cycle; water-phlegm and water dampness were the basic pathological products; toxin-heat toxin, water toxin, and stasis toxin were the final results of disease progress and product accumulation. In terms of treatment, CHF can be divided into four stages: early, middle, late and end. In the early stage, tonifying qi and regulating heart can be used for the treatment of root cause, and promoting blood circulation and water can be used for the treatment of symptoms; tonifying qi and yin and reinforcing the healthy qi, reducing blood stasis, purging turbid, and eliminating pathogenic factors can be used in the middle stage; reducing blood stasis and removing toxic materials should be used in the late stage, supplemented with warming yang and increasing urine excretion; astringing yang,generating body fluids, tonifying qi and yang should be used in the end stage. At the same time of treating by stages, attention should be paid to adhering to a holistic concept and dialectical treatment; pay attention to timing and flexible medication; adopting a combination of Chinese and Western approaches and integrating them.

Application of the Mathieu combined tunnel technique for repairing glans dehiscence after failed hypospadias repair / 亚洲男科学杂志(英文版)

Repairing glans dehiscence after failed hypospadias repair is challenging for pediatric surgeons. Here, we introduced and evaluated a newly modified Mathieu technique, Mathieu combined tunnel (MCT), which involves multiple custom-designed flaps for the shortage of flap source material after repeated operations; we also constructed a tunnel to avoid the glans incision that may carry new risks of dehiscence. This retrospective study included 26 patients who were consecutively admitted to the First Affiliated Hospital of Sun Yat-Sen University (Guangzhou, China) for glans dehiscence repair after failed hypospadias repair from October 2014 to October 2020; sixteen patients underwent surgery using the MCT (MCT group) and ten patients underwent surgery using the tubularized incised plate (TIP) technique (TIP group). The operative time, blood loss, postoperative complications, normal urethral meatus rate, success rate, and Hypospadias Objective Penile Evaluation (HOPE) score were compared between the two groups. The MCT group achieved an overall satisfactory penile appearance and voiding function, with a higher rate of normal urethral meatus (15/16, 93.8%) and a lower rate of glans dehiscence (1/16, 6.2%), compared with the TIP group (70.0% and 30.0%, respectively). However, these differences were not statistically significant, possibly because of the limited number of patients (all P > 0.05). Mean postoperative HOPE scores were similar in the MCT group (mean ± standard deviation: 8.83 ± 0. 89) and TIP group (8.94 ± 0.57) (P > 0.05). No significant differences were found between the two groups in terms of blood loss and success rate, nor in the rates of various complications (e.g., fistula, urethral stricture, and glans dehiscence). In conclusion, the MCT technique appears to be feasible and reliable for repairing glans dehiscence after failed hypospadias repair.

Effects of ginkgo biloba tablets on liver injury in patients with coal-burning-borne arsenism based on DNA damage and repair inhibition / 中华地方病学杂志

Objective:To investigate the role of DNA damage and repair inhibition in the effect of ginkgo biloba on liver injury in patients with coal-burning-borne arsenism.Methods:In March 2017, the investigation was conducted in Jiaole village arsenic poisoning area in Yuzhang Town, Xingren County, Guizhou Province. According to the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) and the "Diagnostic Criteria of Occupational Toxic Hepatopathy" (GBZ 59-2010), 52 patients with arsenism were selected as the ginkgo biloba intervention group, and 49 cases of arsenism patients as intervention control group. Ginkgo biloba tablets were given orally for 3 months (1 tablet/time, 3 times/d) according to the commonly used clinical methods, and no other drugs were given to all subjects during the intervention period. The intervention control group was given placebo in the same way as that of ginkgo biloba intervention group. A total of 41 residents who did not burn high arsenic coal 12 km away with no abnormal liver function were selected as normal control group. Physical examinations were performed before the intervention and at the end of the intervention at 3 months. After receiving signed informed consent, morning urine and peripheral venous blood samples were collected to detect urinary arsenic content by inductively coupled plasma mass spectrometry (ICP-MS); liver function biochemical indexes [albumin (ALB), albumin/globulin (A/G), cholinesterase (CHE), total bile acid (TBA)] were determined by automatic biochemical analyzer, DNA damage by single-cell gel electrophoresis assay, and the expression of miR-145 (repair inhibition index) by qRT-PCR.Results:There were 116 subjects, 41 in normal control group, 39 in ginkgo biloba intervention group and 36 in intervention control group. In ginkgo biloba and intervention and intervention control groups, there was no significant difference in age, gender, smoking habits and drinking compared with normal control group ( P > 0.05). Urinary arsenic content, TBA level, DNA damage degree [comet tail DNA percentage (TailDNA%) and olive tail moment (OTM)] and plasma miR-145 expression level [(38.75 ± 19.09) μg/g Cr, (11.13 ± 1.55) μmol/L, 8.50 ± 0.88, 7.43 ± 0.68, 5.78 ± 0.75, respectively] in ginkgo biloba intervention group patients before intervention were higher than those in normal control group [(11.62 ± 5.33) μg/g Cr, (5.36 ± 0.87) μmol/L, 5.24 ± 0.33, 4.71 ± 0.29, 2.05 ± 0.27, respectively], the differences were statistically significant ( P < 0.05); the levels of ALB, A/G and CHE were significantly lower than those in normal control group ( P < 0.05). After the intervention of ginkgo biloba, urinary arsenic content, TBA level, DNA damage degree (TailDNA% and OTM) and plasma miR-145 expression level in patients were significantly lower than those before the intervention ( P < 0.05); the levels of ALB, A/G and CHE were significantly higher than those before the intervention ( P < 0.05). There was no significant difference in the above indexes before and after intervention in the intervention control group ( P > 0.05). The results of correlation analysis between DNA damage degree, miR-145 and liver function indexes after the intervention of ginkgo biloba showed that, DNA damage degree (TailDNA% and OTM) was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.34, - 0.33, - 0.48, - 0.31, - 0.31, - 0.42, P < 0.05), and positively correlated with the level of TBA ( r = 0.49, 0.48, P < 0.05); miR-145 was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.26, - 0.23, - 0.38, P < 0.05), which was positively correlated with the level of TBA ( r = 0.32, P < 0.05); and DNA damage degree was positively correlated with the expression of miR-145 ( r = 0.65, 0.52, P < 0.05). Conclusion:Ginkgo biloba tablets can alleviate the liver damage caused by arsenic through coal burning, and the mechanism of this process is related to its inhibition of miR-145 expression and reduction of DNA damage.

Application value of right minimal invasive three-port technique of laparoscopic sleeve gastrec-tomy for the treatment of obesity / 中华消化外科杂志

Objective:To investigate the application value of right minimal invasive three-port technique of laparoscopic sleeve gastrectomy (RMIT-LSG) for the treatment of obesity.Methods:The retrospective and descriptive study was conducted. The clinical data of 66 obesity patients who underwent RMIT-LSG in the Sir Run Run Shaw Hospital of Zhejiang University School of Medicine from January to October 2021 were collected. There were 15 males and 51 females, aged 28.5(range, 16.0?54.0)years. The body mass index (BMI) of the 66 patients was (36.9±4.3)kg/m 2. There were 20 of the 66 patients combined with type 2 diabetes. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination or the WeChat to detect postoperative recovery of patients including body mass changing, BMI and complications 6 months after operation. The follow-up was up to December 2021. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical situations. All the 66 patients underwent RMIT-LSG successfully, without conversion to laparotomy or changing surgical method. The operation time and the volume of intraoperative blood loss of the 66 patients were (132±22)minutes and (14±8)mL, respectively. (2) Postoperative situations. The time to postoperative initial out-of-bed activities, time to postoperative first flatus, time to postoperative initial water intake, time to postoperative initial liquid food intake and duration of postoperative hospital stay of the 66 patients were (15±6)hours, (1.80±0.60)days, (1.00±0.20)days, (2.00±0.20)days and (3.40±0.60)days, respectively. Of the 66 patients, one case underwent post-operative abdominal hemorrhage at postoperative day 1 and received a second surgery for hemostasis. The patient with postoperative abdominal hemorrhage and other 65 patients recovered well without gastroparesis, gastric fistula, abdominal infection and other complication. (3) Follow-up. All the 66 patients were followed up for 6(range, 1?11)months. All the 66 patients completed the postoperative scar photography at postoperative 1 month, and results of scar photography showed concealed scar with good cosmetic effects. Twenty-seven of the 66 patients were followed up for 6 months after operation, with the weight loss, percentage of weight loss and decrease of BMI were (42±7)kg, 34.8%±2.9%, (14.2±1.9)kg/m 2, respectively. None of the 66 patient had innutrition during the follow-up. Conclusion:The RMIT-LSG is safe and feasible for the treatment of obesity, with a good cosmetic effect of the wound.

Micro RNA-373-3p is involved in regulation of autophagy and sunitinib sensitivity of glioblastoma cells via targeting autophagy-related gene 7 / 中华神经医学杂志

Objective:To investigate the influence and mechanism of micro RNA (miR)-373-3p in autophagy and sunitinib sensitivity of glioblastoma cells.Methods:U251 cells were routinely cultured in vitro; and some U251 cells were subjected to 50 μmol/L sunitinib treatment for 72 h to construct sunitinib-resistant U251 cell line. (1) Real-time reverse transcription quantitative PCR (RT-qPCR) was used to detect the miR-373-3p expression in U251 and sunitinib-resistant U251 cells. Sunitinib-resistant U251 cells were divided into blank control group, nonsense sequence group and miR-373-3p mimic group; cells in the latter 2 groups were transfected with nonsense sequence and miRNA-337-3p mimic, respectively; miR-373-3p expression was detected by RT-qPCR. Cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, and sunitinib-resistant U251+miR-373-3p mimic group; after each transfection, CCK-8 assay was used to evaluate the cell viability; TUNEL was used to detect the apoptotic rate; immunofluorescent assay was used to detect the expression of microtubule-associated protein light chain 3 (LC3); Western blotting was used to detect the expressions of apoptosis- and autophagy-associated proteins. (2) The pGL3-autophagy-related gene 7 (ATG7) wild-type (WT) and pGL3-ATG7 mutant type (MUT) plasmids were established; dual-luciferase reporter system was used to detect the cell luciferase activity in the miR-373-3p mimic group and nonsense sequence group. Cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, and sunitinib-resistant U251+miR-373-3p mimic group; after each transfection, RT-qPCR and Western blotting were used to detect the mRNA and protein expressions of ATG7 in the cells. (3) The sunitinib-resistant U251 cells were divided into blank control group, ATG7 negative control group, and ATG7 overexpression group; after each transfection, RT-qPCR and Western blotting were used to detect the ATG7 mRNA and protein expressions. U251 and sunitinib-resistant U251 cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, sunitinib-resistant U251+miR-373-3p mimic group, sunitinib-resistant U251+miR-373-3p mimic+ATG7 negative control group, and sunitinib-resistant U251+miR-373-3p mimic+ATG7 overexpression group; after each transfection, CCK-8 assay was used to evaluate the cell apoptosis, TUNEL was used to examine the apoptotic rate, and Western blotting was employed to detect the expressions of apoptosis- and autophagy-associated proteins. Results:(1) As compared with that in the U251 cells, miR-373-3p was lowly expressed in sunitinib-resistant U251 cells, with statistic difference ( P<0.05). As compared with that in the blank control group and nonsense sequence group, miR-373-3p expression was significantly elevated in the miR-373-3p mimic group ( P<0.05). As compared with the U251 group, the sunitinib-resistant U251 group had significantly increased cell viability, significantly decreased cell apoptotic rate, statistically increased B lymphocytoma-2 (Bcl-2) and Beclin 1 protein expressions, and significantly increased LC3II/LC3I values, significantly decreased Bcl-2 associated X protein (Bax) and p62 protein expressions and cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had significantly decreased cell viability, significantly increased cell apoptotic rate, statistically decreased Bcl-2 and Beclin 1 protein expressions, and significantly decreased LC3II/LC3I values, significantly increased Bax and p62 protein expressions and cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the U251 group, the sunitinib-resistant U251 group had increased number of fluorescent particles labeled with LC3 and enhanced fluorescent intensity; as compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had decreased number of fluorescent particles labeled with LC3 and reduced fluorescent intensity. (2) The luciferase activity of pGL3-ATG7 WT plasmids in the miR-373-3p mimic group was signficantly lower than that in nonsense sequence group ( P<0.05). As compared with those in the U251 group, ATG7 mRNA and protein expressions were both significantly increased in the sunitinib-resistant U251 group ( P<0.05); as compared with those in the sunitinib-resistant U251+nonsense sequence group, ATG7 mRNA and protein expressions were both significantly decreased in the sunitinib-resistant U251+miR-373-3p mimic group ( P<0.05). (3) As compared with the blank control group and ATG7 negative control group, the ATG7 overexpression group had significantly increased ATG7 mRNA and protein expressions ( P<0.05). As compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had significantly decreased cell viability, significantly increased cell apoptotic rate, statistically decreased Bcl-2 and Beclin 1 protein expressions, significantly decreased LC3II/LC3I values, significantly increased Bax and p62 protein expressions, and significantly increased cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the sunitinib-resistant U251+miR-373-3p mimic+ATG7 negative control group, the sunitinib-resistant U251+miR-373-3p mimic+ATG7 overexpression group had significantly increased cell viability, significantly decreased apoptotic rate, statistically increased Bcl-2 and Beclin 1 protein expressions, significantly increased LC3II/LC3I values, significantly decreased Bax and p62 protein expressions, and significantly decreased cleaved Caspase3/Caspase 3 values ( P<0.05) Conclusion:MiR-373-3p can enhance sunitinib sensitivity by regulating autophagy in glioblastoma cells, whose mechanism might be related to targeting ATG7.

Performance of loop-mediated isothermal amplification (LAMP) assay for detection of Schistosoma japonicum infection in Oncomelania snails in schistosomiasis transmission-interrupted regions / 中国血吸虫病防治杂志

Objective To compare the effectiveness of loop-mediated isothermal amplification (LAMP) assay and microscopic examinations for detection of Schistosoma japonicum infections in Oncomelania hupensis in transmission-interrupted regions, so as to provide insights into the optimization of snail surveillance tools in these regions. Methods Four hilly schistosomiasis-endemic villages where transmission interruption was achieved were selected in Heqing County of Yunnan Province as the study villages, including Xinzhuang and Gule villages in hilly regions and Lianyi and Yitou villages in dam regions. Snail survey was performed by means of systematic sampling combined with environmental sampling in July 2018. All captured snails were identified for S. japonicum infections using microscopy. In addition, 10 to 20 snails were randomly sampled from each snail habitat following microscopy, numbered according to environments and subjected to LAMP assay. The positive rate of settings with S. japonicum-infected snails was compared among villages. Results A total of 7 949 living snails were captured from 83 snail habitats in 4 villages, and no S. japonicum infection was detected in snails. There were 226 mixed samples containing 1 786 snails subjected to LAMP assay, and positive LAMP assay was found in 3 mixed samples from 3 snail habitats in 2 dam villages. The positive rates of settings with S. japonicum-infected snails were comparable between Lianyi Village (one setting) and Yitou Village (2 set tings) (5.89% vs. 14.29%, P = 0.344). However, the overall positive rate of settings with S. japonicum-infected snails was significantly higher in dam villages (9.67%, 3/31) than in hilly villages (0) (P = 0.048). Conclusions LAMP assay is more sensitive to detect S. japonicum infections in O. hupensis than conventional microcopy method, which may serve as a supplementary method for detection of S. japonicum infections in O. hupensis in high-risk snail habitats in hilly transmission-interrupted regions.

Improved synthetic process of apremilast / 中国药科大学学报

@#To optimize the process of hydrogenation reduction in the synthesis of apremilast (APST), 3-nitrophthalic anhydride (4) and (S)-1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethanamine-(S)-2-acetamido-4-methylpentanoate (7) were used as starting materials to synthesize (S)-2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl)-4-nitroisoindoline-1,3-dione (8) by amination.Then compound 8 was reduced to (S)-4-amino-2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethyl) isoindoline-1,3-dione (9) with ammonium formate as hydrogen source and palladium hydroxide as catalyst.Finally, apremilast was obtained by the acetylation reaction with acetic anhydride.The structure of the products were verified by optical rotation, 1H NMR, 13C NMR, MS and elemental analysis.And the total yield of three steps was increased to 67.0%.The improved reduction process can avoid the special reaction of hydrogenation and pressurization, and reduce the safety risk and production costs with high commercial value.

Classification Model of Corneal Opacity Based on Digital Image Features / 中国医疗器械杂志

OBJECTIVE@#According to the digital image features of corneal opacity, a multi classification model of support vector machine (SVM) was established to explore the objective quantification method of corneal opacity.@*METHODS@#The cornea digital images of dead pigs were collected, part of the color features and texture features were extracted according to the previous experience, and the SVM multi classification model was established. The test results of the model were evaluated by precision, sensitivity and @*RESULTS@#In the classification of corneal opacity, the highest @*CONCLUSIONS@#The SVM multi classification model can classify the degree of corneal opacity.

Effect of dictyophora polysaccharide on mitophagy induced by sodium arsenite in L-02 cells / 中华地方病学杂志

Objective:To observe the effect of dictyophora polysaccharide (DIP) on PINK1/Parkin pathway mediated mitophagy induced by sodium arsenite (NaAsO 2) in human hepatocytes (L-02 cells). Methods:The L-02 cells in logarithmic growth phase and in good condition were divided into control group, NaAsO 2 group (10 μmol/L), DIP group (80 μg/ml), DIP + NaAsO 2 group (80 μg/ml DIP + 10 μmol/L NaAsO 2) , N-acetylcysteine (NAC) group (5 mmol/L), and NAC + NaAsO 2 group (5 mmol/L NAC + 10 μmol/L NaAsO 2). Western blotting was used to detect the expression levels of mitophagy related proteins p62, microtubule-associated protein 1 light chain 3 (LC3)Ⅱ/LC3Ⅰ, PINK1, and Parkin. The mitochondrial stucture and autophagosomes were observed by transmission electron microscope, the fluorescent probe method was used to detect the expression level of intracellular reactive oxygen species (ROS). Results:Compared with the control group, the protein expressions of p62, LC3 Ⅱ/LC3 Ⅰ, PINK1, and Parkin in NaAsO 2 group were higher ( P < 0.05); compared with the NaAsO 2 group, the protein expressions of p62, LC3 Ⅱ/LC3 Ⅰ, PINK1 and Parkin were lower in DIP, DIP + NaAsO 2, NAC, and NAC + NaAsO 2 groups ( P <0.05). According to the transmission electron microscope, compared with the control group, the mitochondria of L-02 cells in NaAsO 2 group were significantly damaged and the number of autophagosomes increased. Compared with NaAsO 2 group, the degree of mitochondrial swelling, vacuolar degeneration and the number of autophagosomes decreased in DIP + NaAsO 2 group. Compared with the control group (33 110.00 ± 2 191.28), the intracellular ROS level in NaAsO 2 group was higher (48 000.00 ± 2 395.31, P < 0.05); the level of intracellular ROS in DIP + NaAsO 2 group (38 670.00 ± 2 620.56) was significantly lower than that in NaAsO 2 group( P < 0.05), and there was no significant change compared with the control group ( P > 0.05). Conclusions:NaAsO 2 can induce PINK1/Parkin mediated mitophagy in L-02 cells. DIP can alleviate NaAsO 2 induced mitophagy. DIP may affect PINK1/Parkin mediated mitophagy induced by NaAsO 2 through the regulation of ROS.

Research progresses of radiomics in thyroid nodules / 中国医学影像技术

Radiomics can extract quantitative features from medical images and perform quantitative evaluation of tumors, so as to assist the diagnosis, treatment and prognosis evaluation, having great potential in diagnosis and treatment of tumors. With the increasing detection rate of thyroid nodules, radiomics has been gradually applied into studies of thyroid nodules. The research progresses of radiomics in thyroid nodules were reviewed in this article.

Role of reactive oxygen species-mitochondrial pathway in sodium arsenite-induced apoptosis of L-02 cells / 中华地方病学杂志

Objective:To explore the mechanism of apoptosis induced by sodium arsenite (NaAsO 2) in human hepatic cells (L-02) through reactive oxygen species (ROS) accumulation and mitochondrial dysfunction, and provide experimental evidence for the mechanism of arsenic poisoning. Methods:L-02 cells were divided into control group, NaAsO 2 group (10 μmol/L NaAsO 2), N-acetylcysteine (NAC) group (5 mmol/L NAC), and NaAsO 2 + NAC group (10 μmol/L NaAsO 2, 5 mmol/L NAC), and were cultured in vitro for 24 h. The intracellular ROS level, mitochondrial membrane potential depolarization ratio and cell apoptosis rate were measured by dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence probe, JC-1 staining and Annexin V-FITC/PI double staining, respectively; the mRNA and the protein of Caspase 3, cytochrome C (Cyt-C) and cytochrome C oxidaseⅣ (COXⅣ) were detected by real time fluorescence quantitative PCR (qRT-PCR) and Western blotting, respectively. Results:There were statistically significant differences in intracellular ROS levels (3 857 392.33 ± 44 928.39, 4 515 288.00 ± 32 660.64, 3 670 150.67 ± 101 987.69, 4 035 235.67 ± 99 995.30), mitochondrial membrane potential depolarization ratios (2.16 ± 0.54, 7.95 ± 0.52, 2.70 ± 0.29, 1.01 ± 0.23) and total apoptosis rates (1.45 ± 0.03, 4.27 ± 0.17, 1.87 ± 0.12, 2.52 ± 0.35) between groups ( F = 62.62, 159.81, 112.70, P < 0.05). There were statistically significant differences in Caspase 3, Cyt-C, COXⅣ mRNA expression levels ( F = 9.20, 7.33, 14.87, P < 0.05) and in cleaved-Caspase 3, Cyt-C, COXⅣ protein expression levels( F = 31.42, 8.01, 83.30, P < 0.05) between groups. Compared with the control group, the intracellular ROS level, mitochondrial membrane potential depolarization ratio and total apoptosis rate were significantly increased ( P < 0.05); Caspase3, Cyt-C mRNA and protein expression levels were significantly increased ( P < 0.05), and COXⅣ mRNA and cleaved-Caspase 3, Cyt-C protein expression levels were significantly decreased ( P < 0.05) in NaAsO 2 group. Compared with the NaAsO 2 group, the intracellular ROS level, mitochondrial membrane potential depolarization ratio and total apoptosis rate of NaAsO 2 + NAC group were significantly decreased ( P < 0.05); the Caspase3, Cyt-C mRNA and cleaved-Caspase 3, Cyt-C protein expression levels were significantly decreased ( P < 0.05), the COX Ⅳ mRNA and protein expression levels were significantly increased ( P < 0.05). Conclusions:NaAsO 2 stimulates L-02 cells to produce excessive ROS, which induces mitochondrial depolarization and further triggers mitochondrial damage, resulting in increased release of Cyt-C and activation of the mitochondrial apoptosis pathway that Caspase 3 protein induces apoptosis in L-02 cells, which may be one of the main mechanisms of arsenic-induced liver injury.

Role of Nrf2 signaling pathway in oxidative damage of L-02 cells induced by sodium arsenite / 中华地方病学杂志

Objective:To explore the role of nuclear factor-E2-related factor 2 (Nrf2) signaling pathway in oxidative damage caused by sodium arsenite (NaAsO 2) in human normal liver cells (L-02), and to provide experimental basis for the study of oxidative damage mechanism of liver damage caused by arsenic. Methods:L-02 cells were cultured in vitro and treated with 0 (control), 25, 50, 75, 100, 125, and 150 μmol/L NaAsO 2, respectively, for 24 h. The half-inhibitory concentration (IC 50) was calculated according to the cell survival rate by CCK8, and L-02 cells were treated with 0, 1/8, 1/4 and 1/2 dose of IC 50 of NaAsO 2, respectively, for grouping experiments. Protein expressions of Nrf2, heme oxygenase-1 (HO-1), NADH quinone oxidoreductase 1 (NQO1) and glutathione peroxidase 1 (GPx1) in L-02 cells and L-02 nucleus were detected by Western blotting. Results:The result of CCK8 showed that the survival rates of L-02 cells in 25, 50, 75, 100, 125, 150 μmol/L NaAsO 2 groups were [(69.53 ± 0.06)%, (41.33 ± 0.08)%, (23.65 ± 0.04)%, (26.51 ± 0.04)%, (31.63 ± 0.01)%, (26.24 ± 0.02)%], which were significantly lower than that of the control group[(100 ± 0.00)%]. The differences were statistically significant ( P < 0.05). The IC 50 calculated by cell survival was 40 μmol/L, and the NaAsO 2 doses used in the experiment were 0 (control), 5, 10, and 20 μmol/L. Western blotting results showed that, compared with the control group, the protein expression levels of Nrf2, HO-1 in L-02 and HO-1 in the L-02 cells nucleus in the 5, 10 and 20 μmol/L NaAsO 2 groups were significantly higher ( P < 0.05). Compared with the control group, the expression levels of GPx1 protein in L-02 cells of 10 and 20 μmol/L NaAsO 2 groups were decreased ( P < 0.05). Compared with the control group, the expression levels of Nrf2 protein in L-02 nucleus in 10 and 20 μmol/L NaAsO 2 groups were significantly increased ( P < 0.05); the expression level of NQO1 protein in L-02 nucleus in 5 μmol/L NaAsO 2 group was significantly increased ( P < 0.05). Conclusion:NaAsO 2 has an effect on the expression of Nrf2 signaling pathway related factors in L-02 cells, and the mechanism of oxidative damage caused by NaAsO 2 in L-02 cells may be related to Nrf2 signaling pathway.

Effects of sodium arsenite on the expression of lipid metabolism related genes SREBP-1c, PPARα and FAS in L-02 cells / 中华地方病学杂志

Objective:To investigate the effects of sodium arsenite (NaAsO 2) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator activated receptor α (PPARα) and fatty acid synthase (FAS) in human liver cells (L-02 cells). Methods:L-02 cells were cultured in vitro, and exposed to NaAsO 2 at 0 (control), 2, 4, 8, 16, 32, 64 and 128 μmol/L for 24 h, respectively, and the cell survival rate was determined by CCK-8 method. And a fitting curve was made to calculate the half inhibitory concentration (IC 50), subsequent experiments were carried out with 0, 1/8, 1/4 and 1/2 of IC 50 as arsenic exposure doses. Glycerol phosphate oxidase-catalase (GPO-PAP) method was used to detect the content of triglyceride (TG) in cells; the mRNA expression levels of SREBP-1c, PPARα and FAS were detected by Real-time PCR; and the protein expression levels of SREBP-1c and PPARα were detected by Western blotting. Results:The cell survival rates of 8, 16, 32, 64 and 128 μmol/L NaAsO 2 groups [(92.000 ± 1.414)%, (91.000 ± 0.000)%, (76.500 ± 0.707)%, (53.000 ± 1.412)%, (47.000 ± 1.412)%] were significantly lower than that of the control group [(100.000 ± 0.000)%, P < 0.01]. The IC 50 was 64 μmol/L, and subsequent experiments were conducted with 0 (control), 8, 16 and 32 μmol/L NaAsO 2, respectively. Compared with the control group [(1.000 ± 0.000) mmol/g prot], TG contents of 8, 16 and 32 μmol/L NaAsO 2 groups [(0.691 ± 0.064), (0.474 ± 0.162), (0.184 ± 0.045) mmol/g prot] were significant decreased ( P < 0.01). Compared with the control group, the mRNA expression levels of SREBP-1c, PPARα, FAS, and the protein expression levels of SREBP-1c and PPARα in NaAsO 2 groups were significantly decreased ( P < 0.01 or < 0.05). Correlation analysis showed that NaAsO 2 content was negatively correlated with TG content, SREBP-1c and PPARα protein expression levels ( r =-0.954,- 0.875,-0.965, P < 0.01). Conclusion:NaAsO 2 can reduce the TG content and the expression of lipid metabolism related genes SREBP-1c, PPARα and FAS in L-02 cells, suggesting that arsenic-induced liver injury can cause lipid metabolism disorders.

Application of teaching with multi-modality visualization in the training of neurosurgical residents / 中华医学教育探索杂志

Objective To teach neurosurgical residents of standardized training by using teaching method with multi-modality visualization and to explore its application effects.Methods Total 122 students were randomly divided into two groups:multi-modality visualization teaching group (n=61) and traditional teaching group (n=61).The evaluation of teaching effect was conducted by questionnaire of students and the analysis of test scores after the course.Comparison between the two groups was made by using independent sample t test.Results Questionnaire showed that the satisfaction of teaching mode (88.5％),learning efficiency (93.4％),and training results (90.1％) with multi-modality visualization teaching group were statistically higher than traditional group (P＜0.05).Test score showed that results of theory test (88.5 ± 5.1),on-spot examination (91.6 ± 5.5),and overall score (89.3 ± 5.2) were also statistically higher in multi-modality visualization teaching group than that of control group (P＜0.05).There was no significant difference in clinical skills assessment between two groups.Conclusions Teaching with multi-modality visualization can significantly improve the efficiency of neurosurgical clinical teaching and promote the training effect of students,which provides a new strategy of neurosurgical clinical teaching.

Effects of three MAPK inhibitors on the expressions of TGF-β1 and α-SMA mRNA and protein in LX-2 cells induced by sodium arsenite / 中华地方病学杂志

Objective To investigate the effects of three mitogen-activated protein kinase (MAPK) inhibitors on the expressions of transforming growth factor-β1 (TGF-β1),α-smooth actin (α-SMA) mRNA and protein in human liver stellate cells (LX-2 cells) activated by sodium arsenite.Methods Cultured in vitro LX-2 cells in the logarithmic growth stage were exposed to sodium arsenite at 0.0 (control),2.5,5.0,10.0,20.0,40.0,80.0 μmol/L for 24 h,respectively,and the cell survival rate was determined by CCK-8 assay.According to the results of the study,LX-2 cells were divided into 5 groups:control group,sodium arsenite group,extracellular signal regulation kinase (ERK) inhibition group,c-Jun amino-terminal kinase (JNK) inhibition group,and p38 inhibition group.LX-2 cells were pre-treated with 10.0 μmol/L ERK,JNK,p38 kinase inhibitors (PD98059,SP600125,SB203580) for 30 min in the 3 inhibition groups,and then 20.0 μmol/L sodium arsenite for 24 h.The control group was not treated with sodium arsenite and inhibitors.Sodium arsenite group was not treated with inhibitors.Then mRNA and protein expression levels of TGF-β1 and α-SMA in LX-2 cells were determined by Western blotting and real-time PCR,respectively.Results The survival rates of LX-2 cells in 5.0,10.0,20.0,40.0,80.0 μmol/L sodium arsenite groups were [(92.35 ± 0.92)％,(84.06 ± 0.84)％,(74.27 ± 0.74)％,(59.57 ± 0.60)％,(27.77 ± 0.23)％],which were significantly lower than that of the control group [(100.00 ± 0.00)％,P ＜ 0.05].It was found that the expressions of TGF-β1,o-SMA mRNA and protein of sodium arsenite group were higher than those of the control group (P ＜ 0.01).The expressions of TGF-β1,α-SMA mRNA and protein of the three inhibition groups were lower than those of the sodium arsenite group (P ＜ 0.05).Conclusions Arsenic exposure can cause abnormally high expressions of TGF-β1,α-SMA mRNA and protein in LX-2 cells.Intervention with three MAPK inhibitors can improve the effects of arsenic induced LX-2 cells activation on the expressions of TGF-β1,α-SMA mRNA and protein.