Mitochondria play a central role in the regulation of energy metabolism and signal transduction in eukaryotic cells. Although many fluorescent labeling strategies have been developed for mitochondrial studies, the methods that enable labeling efficiency assessment at the single-mitochondrion level are still lacking. By employing the unique advantages of high sensitivity flow cytometry ( HSFCM ) in the sensitive, rapid, and quantitative multiparameter analysis of individual mitochondria, here we examined the performance of several different mitochondrial labeling strategies from the perspectives of brightness, labeling ratio, and stability. Mitochondria isolated from HeLa cells transfected with pAcGFP1-Mito plasmid upon transient or stable transfections, and mitochondria directly labeled with MitoTracker Green or SYTO 62 were analyzed by a laboratory-built three-channel HSFCM. Upon the quantitative measurement of fluorescence brightness, it was found that the fluorescence intensity of green fluorescent protein ( GFP ) in mitochondria isolated from cells with stable transfection was about 17. 7-fold higher than the transient transfection ones, and was approximately two orders of magnitude brighter than mitochondria labeled with MitoTracker Green. On the other hand, the fluorescence signal of SYTO 62 labeling decreased upon washing, indicating its rapid dissociation rate. The strong fluorescence intensity and good labeling stability make stable transfection an efficient method to label mitochondria. The experimental results demonstrates that HSFCM provides a powerful analytical tool to assess the performance of mitochondrial fluorescence labeling via high throughput single mitochondria analysis.

Objective:To evaluate the effect of dexmedetomidine on the quality of recovery from anesthesia in elderly patients undergoing electroconvulsive therapy (ECT) with propofol anesthesia.Methods:Sixty patients of both sexes, aged>65 yr, weighing 45-80 kg, of American Society of Anesthesiologists physical status Ⅰor Ⅱ, scheduled for elective ECT with propofol anesthesia, were assigned into 2 groups ( n=30 each) using a random number table method: control group (group C) and dexmedetomidine group (group D). Dexmedetomidine was intravenously infused in a dose of 0.2 μg/kg (in normal saline 10 ml) over 10 min starting from onset of anesthesia induction in group D, while normal saline 10 ml was given instead in group C. Propofol 1.0-1.5 mg/kg was intravenously injected slowly.Succinylcholine 0.7 mg/kg was intravenously injected after the eyelash reflex disappeared, and oxygen was delivered via a mask to assist artificial ventilation.The mask was removed when the muscle twitching disappeared during depolarization, treatment was performed with an ETC apparatus, and electroencephalogram was monitored.The electrical stimulus intensity was set according to the age of the patient during ECT treatment, and the initial intensity was set at 0.5 times the age of the patient. When the postictal suppression index was less than 80%, a higher level of stimulus intensity was used in the next ECT treatment (the difference between adjacent intensity levels was 25.2 mc, which was 5% of the total stimulus intensity). After the end of ECT procedure, participants were manually ventilated with a mask, and the patients were transferred to postanesthesia care unit when the spontaneous breathing was completely restored.The time to recovery of spontaneous breathing and emergence time were recorded.The development of adverse cardiovascular events, nausea and vomiting, respiratory depression, headache, drowsiness, agitation and delirium during recovery from anesthesia was also recorded. Results:Compared with group C, the incidence of agitation, delirium, hypertension and tachycardia during recovery from anesthesia was significantly decreased, and no significant change was found in the other variables in group D ( P<0.05). Conclusion:Dexmedetomidine can improve the quality of recovery from anesthesia in elderly patients undergoing ECT under propofol anesthesia.

Objective:In recent years,the prevalence of diabetic nephropathy(DN)has increased significantly.An increasing number of studies have shown that lymphocyte-associated inflammatory responses play a role in DN.This study aims to investigate the relationship between lymphocytes and DN in patients with autoimmune diabetes. Methods:The clinical data of 226 patients with Type 1 diabetes(T1D)and 79 patients with latent autoimmune diabetes in adults(LADA)were retrospectively studied and stratified according to the urinary albumin to creatinine ratio(ACR).Risk factors associated with DN were analyzed using correlation analysis and logistic regression. Results:In T1D and LADA patients,systolic blood pressure(SBP),uric acid duration,and diabetes duration in patients with normoalbuminuria were lower or shorter than those in patients with macroalbuminuria(P＜0.05).The lymphocyte count of T1D patients was significantly higher than that in LADA patients(P＜0.05),while the neutrophil to lymphocyte ratio(NLR)of T1D patients was significantly lower than that in LADA patients(P＜0.05).The lymphocyte count in the T1D patients with normoalbuminuria was lower than that those with macroalbuminuria(P＜0.05).The NLR was lower in the T1D patients with macroalbuminuria than those with microalbuminuria and normoproteinuria(all P＜0.01).Based on logistic regression analysis,lymphocytes were independently associated with DN in T1D after adjusting for various known risk factors such as course of disease,age,gender,dyslipidemia,hypertension,and smoking status.Analysis of the receiver operating characteristic curve of subjects predicting lymphocytes in normoalbuminuria showed that the area under the curve was 0.601(95% CI 0.510 to 0.693,P=0.039),and when the cutoff value of lymphocytes was 2.332,the sensitivity was 37.0%,and the specificity was 82.5%. Conclusion:Lymphocyte counts in autoimmune diabetic patients are closely associated with DN,suggesting that lymphocyte-mediated inflammation may be involved in the pathogenesis of DN in autoimmune diabetic patients.This study provides a possible perspective for using lymphocytes as a potential biomarker for the early identification of individuals at risk for DN and potential therapeutic targets for DN.

Radiotherapy is a first-line treatment for a variety of malignant tumors by inducing DNA damage to kill tumor cells. However, tumor cells have different sensitivities to radiotherapy, ultimately leading to different therapeutic effects. Histone acetylation, regulated by histone acetyltransferase (HAT) and histone deacetylase (HDAC), is involved in the regulation of cell radiation sensitivity by influencing DNA damage repair. The main mechanisms are recruiting DNA repair related proteins and mediating chromatin dynamic changes. In this article, the role of histone acetylation modification in tumor radiotherapy was reviewed, aming to provide the basis for the radiotherapy sensitization strategy based on histone acetylation.

BACKGROUND:Intervertebral disc degeneration is an important cause of low back pain.At present,there are many modeling methods for disc degeneration in China and abroad,but there is not a model for low back pain due to disc degeneration. OBJECTIVE:To compare the effect of mechanical puncture combined with tumor necrosis factor α and complete Freund's adjuvant with a conventional disc mechanical puncture alone. METHODS:A total of 18 male adult Sprague-Dawley rats were randomly divided into 3 groups,with 6 animals in each group.No treatment was given in the blank group.Animal models of intervertebral disc degeneration were made in the L4-5 segments of rats in the control using conventional mechanical puncture.In the experimental group,on the basis of mechanical puncture,tumor necrosis factor α+complete Freund's adjuvant was injected into the L4-5 intervertebral discs using a microinjector to establish a model of disc degeneration induced by mechanical puncture combined with inflammatory factors.Four weeks after surgery,the pain threshold of rats was measured by the hot plate method for assessing the perception of heat injury in rats with intervertebral disc degeneration.MRI examination was performed to observe the disc degeneration in each group.ELISA was used to detect the levels of serum tumor necrosis factor α,interleukin 1β,interleukin 6 and prostaglandin E2.Hematoxylin-eosin and Safranin O-fast green staining were used to observe the morphological changes of the disc. RESULTS AND CONCLUSION:In terms of pain,the behavioral pain threshold of the experimental group was continuously decreased,and the levels of serum inflammatory factors were significantly higher compared with the control group.In terms of morphology,the MRI results showed that the L4-5 nucleus pulposus signal completely disappeared in the experimental group.Histopathological results showed that in the control group,the nucleus pulposus was intact,more notochord cells were visible,and some fiber rings were ruptured,while in the experimental group,there are fewer notochord cells and the structure of the nucleus pulposus and fibrous ring is disturbed,with the boundary disappearing.To conclude,mechanical puncture combined with tumor necrosis factor alpha and complete Freund's adjuvant can successfully establish a discogenic low back pain model in rats.This operation is simple and economical to achieve obvious disc degeneration and low back pain,with greatly shortened molding cycle.This model can be used as a reference for studying discogenic low back pain models.

Objective:To study the expression and clinical significance of teratocarcinoma-derived growth factor 1 (Cripto-1) and mammalian target of rapamycin (mTOR)in cervical cancer (CC)cancer tissues.Methods:From January 2012 to May 2017, 152 patients with CC in the 2nd Affiliated Hospital of Chengdu Medical College were selected as CC group, and 40 patients with uterine fibroids as control group. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Cripto-1 and mTOR in 152 cases of CC tissues and 40 cases of normal cervical tissues. The expression difference between the two and their relationship with pathological features and prognosis were statistically analyzed.Results:The relative expression of Cripto-1 and mTOR in CC tissues was significantly higher than that in normal cervical tissues ( P<0.05). There was a significant positive correlation between Cripto-1 and mTOR expression in CC tissues ( r=0.634, P<0.05). The expression of Cripto-1 and mTOR protein in CC tissues were associated with International Federation of Gynecology and Obstetrics (FIGO) stage and tumor grade ( P<0.05), but not with age, pathological type, depth of invasion and lymph node metastasis ( P>0.05). The 3-year overall survival (OS) rate of patients with high Cripto-1 expression was not significantly different from that of patients with low Cripto-1 expression ( P>0.05), while the 3-year OS of high mTOR expression group was significantly lower than that of low mTOR expression group (χ 2=5.808, P=0.016). Conclusions:The expression of Cripto-1 and mTOR is increased in CC tissues.Both of them are related to FIGO stage and tumor grade, which may become a new molecular marker for diagnosis, treatment and evaluation of CC.

Purpose: Combining targeted agents with adjuvant chemotherapy prolongs survival in human epidermal growth factor receptor 2 (HER2)-positive breast cancer patients, but also increases the risk of adverse effects. The updated results of 3 randomized controlled trials (RCTs) were reported in 2019. Given the lack of adequate head-to-head pairwise assessment for anti-HER2 agents, network meta-analysis facilitates obtaining more precise inference for evidence-based therapy. Methods: RCTs comparing at least 2 anti-HER2 regimens in an adjuvant setting for HER2-positive early-stage breast cancer (EBC) were included. Hazard ratios for overall survival (OS) and disease free survival (DFS), with respective 95% confidence intervals were pooled for assessment of efficacy. A Bayesian statistical model was used, and odds ratios (ORs) for adverse events (AEs) were used to pool effect sizes. Results: We demonstrated that 1-year trastuzumab plus chemotherapy had increased efficacy compared to shorter or longer treatment duration. The OR of cardiac events gradually increased from 6 months to 1 and 2-year trastuzumab arms, relative to chemotherapy only.Compared to trastuzumab plus chemotherapy, dual HER2-targeting therapies increased DFS, especially for hormone receptor negative patients. Dual anti-HER2 blockade regimens revealed an increased probability of gastrointestinal reactions. As a second agent, pertuzumab showed significantly higher DFS and OS. Conclusion We conclude that 1-year adjuvant trastuzumab should remain as the standard treatment for HER2-positive EBC patients, as it has greater efficacy and a manageable proportion of AEs. Clinical efficacy can be increased for hormone receptor-negative tumors by including a second HER2-targeted agent to the treatment regimen. For hormone receptorpositive cases with basal disease, it is acceptable to reduce the risk of cardiotoxicity by shortening the duration of trastuzumab.

Purpose/Significance To understand the current status and influencing factors of residents'utilization and dissemination of online dietary health information in the Yangtze River Delta region,so as to provide theoretical references for strengthening education on online dietary health information,and eliminating misinformation related to online dietary health.Method/Process The purposive sam-pling method is used to investigate residents in the Yangtze River Delta region,and the data is analyzed by descriptive analysis,inde-pendent sample t-tests,and multiple linear regression analysis.Result/Conclusion The dissemination of online dietary health informa-tion presents interactive characteristics of interpersonal communication and online communication.WeChat and Sina Weibo are the main sources of dietary health information.In addition to information screening,information search,systematic information processing,demand degree,information sharing intention and information concern motivation have significant and positive effects on the public's willingness to receive information.

Objective To verify the safety and efficacy of IONTRIS particle therapy system ( IONTRIS) in clinical implementation. Methods Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial:31 males and 4 females with a median age of 69 yrs ( range 39?80) . Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non?metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results Twenty?two patients received carbon ion and 13 had proton irradiation. With a median follow?up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression?free survival rate was 93.8％ (15/16). Among the 19 patients with prostate cancer, biological?recurrence free survival was 100％. Particle therapy was well tolerated in all 35 patients. Twenty?five patients (71.4％) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow?up. Six ( 17.1％) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow?up is needed to observe the late toxicity and long term result.

Objective To verify the safety and efficacy of IONTRIS particle therapy system ( IONTRIS) in clinical implementation. Methods Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial:31 males and 4 females with a median age of 69 yrs ( range 39?80) . Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non?metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results Twenty?two patients received carbon ion and 13 had proton irradiation. With a median follow?up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression?free survival rate was 93.8％ (15/16). Among the 19 patients with prostate cancer, biological?recurrence free survival was 100％. Particle therapy was well tolerated in all 35 patients. Twenty?five patients (71.4％) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow?up. Six ( 17.1％) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow?up is needed to observe the late toxicity and long term result.