BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

Objective To explore the optimal method for preparing fresh and fixed skeletal muscle tissues,and to lay an experimental foundation for the rapid diagnosis of and research into the pathogenesis of skeletal muscle diseases.Methods The tibialis anterior muscle was extracted from C57BL/6J mice.Fresh tissue was treated by direct rapid freezing in liquid nitrogen,embedding combined with liquid nitrogen freezing,and foreign body alkane treatment combined with liquid nitrogen freezing.Fixed tissues were pre-treated by direct embedding with embedding agent combined with rapid liquid nitrogen freezing.The frozen sections were stained with hematoxylin and eosin.The cross-sectional areas of ice crystals and muscle fibers were calculated to evaluate the effects of the different pre-treatment method.Results The morphology of the muscle fiber bundles was disrupted and numerous ice crystal vacuoles were observed in fresh tissues after direct liquid nitrogen freezing and foreign body alkane treatment combined with liquid nitrogen freezing.In contrast,the muscle fiber bundles were intact and dense and there were no ice crystals in tissues treated with embedding agent combined with rapid liquid nitrogen freezing,indicating that this pre-treatment method was suitable for preparing fresh skeletal muscle tissue.Fixed tissue treated with embedding agent and liquid nitrogen freezing also showed complete muscle fiber bundles and no ice crystals.Conclusions Treatment of fresh and fixed skeletal muscle tissues with embedding agent combined with rapid liquid nitrogen freezing preserves muscle fiber bundles,with no ice crystals.Tissues prepared by this method are thus suitable for further examinations,such as immunohistochemistry and immunofluorescence.This method will therefore aid the accurate and rapid diagnosis of and research into the pathogenesis of skeletal muscle diseases.

Two female patients with acute onset of sporadic food-borne botulism are reported. Both patients presented cranial nerve palsy symptoms and rapidly involving respiratory muscles causing respiratory failure. The tracheal intubation, mechanical ventilation and active supportive measures were given at admission. The skull imaging and cerebrospinal fluid examination were negative, and the electromyography showed peripheral motor nerve damage. One patient was complicated with severe pneumonia. The botulinum toxin test was positive, and botulism poisoning was diagnosed. After early targeted treatment, patients were improved and discharged. Both patients had no complications and recurrence after discharge in 8 months of follow-up.

Objective:To evaluate the prognosis and determine the failure patterns after radiotherapy for low-risk early-stage patients with extranodal NK/T-cell lymphoma, nasal-type (ENKTCL).Methods:A total of 557 patients from 2000—2015 with low-risk early-stage ENKTCL who received radiotherapy (RT) with or without chemotherapy (CT) from China Lymphoma Collaborative Group were retrospectively reviewed. Among them, 427 patients received combined modality therapy, whereas 130 patients received RT alone. Survivals were calculated by Kaplan-Meier method and compared with Log-rank test. Overall survival (OS) was compared with age and sex-matched general Chinese population using expected survival and standardized mortality ratio (SMR). Cox stepwise regression model was used for multivariate analysis.Results:The 5-year OS and progression-free survival (PFS) were 87.2% and 77.2%. The SMR was 3.59 ( P<0.001) at 1 year after treatment, whereas it was 1.50 at 4 years after treatment, without significant difference between ENKTCL group and country-matched general population ( P=0.146). Compared with RT alone, CMT did not result in significantly superior 5-year OS (87.0% vs 87.4%, P=0.961) or PFS (76.1% vs 80.7%, P=0.129). Local failure (11.5%, 64/557) and distant failure (10.8%, 60/557) were the main failure modes, while regional failure was rare (2.9%, 16/557). The 5-year locoregional control rate (LRC) was 87.2% for the whole group, with 89.5% for ≥50 Gy versus 73.7% for <50 Gy ( P<0.001). Radiotherapy dose was an independent factor affecting LRC( P<0.05). Conclusions:Radiotherapy achieves a favorable prognosis in patients with low-risk early-stage ENKTCL. The incidence of either locoregional or distant failure is low. Radiation dose still is an important prognostic factor for LRC.

Objective:To analyze the influencing factors of HIV-testing awareness of Myanmar people in Dehong, and provide references for early discovery of HIV infection among Myanmar people.Methods:A cross-sectional study was conducted among 400 Myanmar people in Dehong through convenient sampling method in a form of face-to-face questionnaire interview. HIV-testing awareness and influencing factors were described and analyzed with χ2 and logistic regression method . Results:A total of 393 participants were included in the study, in whom 241 (61.3%) were males, 256(65.1%) were Myanmar people, 233(59.3%) received 1-7 years of education in Myanmar, 348 do not speak Chinese (88.5%), 226(57.5%) were married, 378(96.2%) were non-solitary, 257(65.4%) were workers, the main purpose of etering into China in 273(69.5%) was working in China, 273(69.5%) had stayed in China for more than 2 years, 573(95.4%) had a clear plan on whether to stay in China for a long time in the future. In these Myanmar people, the rate of AIDS-related knowledge awareness was 75.6%; 165(42.0%) had sexual behavior in the last 6 months. 221(56.2%) knew that they had been tested for HIV. Ninety (22.9%) were more likely to actively seek HIV-testing after they had unprotected behaviors such as homosexual sex, injecting drug use, and sex without using condom. Multivariate logistic regression analysis showed that the influencing factors of HIV-testing awareness included being women (compared with men, OR=2.155, 95% CI: 1.088-4.270), educational level of 8-10 years or more than 10 years (compared with education level 1-4 years, OR=5.207, 95% CI: 2.094-12.950; OR=19.780, 95% CI: 5.800-67.457), having sexual behavior in the last 6 months (compared with those who had not, OR=2.534, 95% CI: 1.343-4.779), having received AIDS-related education in the last 6 months (compared with those who had not, OR=2.462, 95% CI: 1.303-4.654), AIDS-related knowledge awareness (compared with those who had no awareness, OR=8.126, 95% CI: 2.936-22.491). Conclusions:Myanmar people have low awareness of HIV-testing in Dehong. Those who were women and high literacy levels, having received AIDS-related knowledge education in the past 6 months, and having AIDS-related knowledge have an active HIV-testing awareness, after they have high-risk behaviors. AIDS-related knowledge should be publicized to promote early discovery of high-risk behaviors.

Objective:To summarize the experience of radiation oncology residency training programs in western coutries, and provide evidence to improve the present Phase-I Radiation Oncology Residency Training Programs in China.Methods:An electronic questionnaire-based survey was conducted among residents and staffs in 6 top cancer centers in China and 5 centers from Europe and North America to collect their feedback regarding the description and comparison of different programs.Results:A total of 70 responses and 4 papers explaining relevant training programs from 26 residents and 23 teachers in Chinese hopsitals and 20 residents and 1 teacher from Europe and North America were received. The Radiation Oncology Residency Training Programs in China were designed into 2 phases, and the results in the current study were involved with the first phase. Program designs were similar in the following aspects: goal, clinical practice-based training, rotation curriculum, interim and final assessment. However, the total timeframe in the investigated Chinese Phase-I programs was shorter than that in western hospitals (2 to 3 years vs. 4 to 5 years). Chinese programs covered major common diseases, whereas rotation design was performed based on each individual disease in western programs. In Chinese programs, the working hours were mainly 40-60 h every week. Although the working hours were commensurate with local workforce regulations, the residents from MSK program had an outstanding longer working time of 60-80 h every week and treated more patients compared with other programs. Conclusions:The investigated Phase-I Radiation Oncology Residency Training Programs in 6 top cancer centers in China share common features in goals, training modes and assessments with western programs. However, our programs have shorter timeframe and less detailed requirements in individual disease than the western programs.

Objective:To evaluate the prognosis and determine the failure patterns after radiotherapy for low-risk early-stage patients with extranodal NK/T-cell lymphoma, nasal-type (ENKTCL).Methods:A total of 557 patients from 2000—2015 with low-risk early-stage ENKTCL who received radiotherapy (RT) with or without chemotherapy (CT) from China Lymphoma Collaborative Group were retrospectively reviewed. Among them, 427 patients received combined modality therapy, whereas 130 patients received RT alone. Survivals were calculated by Kaplan-Meier method and compared with Log-rank test. Overall survival (OS) was compared with age and sex-matched general Chinese population using expected survival and standardized mortality ratio (SMR). Cox stepwise regression model was used for multivariate analysis.Results:The 5-year OS and progression-free survival (PFS) were 87.2% and 77.2%. The SMR was 3.59 ( P<0.001) at 1 year after treatment, whereas it was 1.50 at 4 years after treatment, without significant difference between ENKTCL group and country-matched general population ( P=0.146). Compared with RT alone, CMT did not result in significantly superior 5-year OS (87.0% vs 87.4%, P=0.961) or PFS (76.1% vs 80.7%, P=0.129). Local failure (11.5%, 64/557) and distant failure (10.8%, 60/557) were the main failure modes, while regional failure was rare (2.9%, 16/557). The 5-year locoregional control rate (LRC) was 87.2% for the whole group, with 89.5% for ≥50 Gy versus 73.7% for <50 Gy ( P<0.001). Radiotherapy dose was an independent factor affecting LRC( P<0.05). Conclusions:Radiotherapy achieves a favorable prognosis in patients with low-risk early-stage ENKTCL. The incidence of either locoregional or distant failure is low. Radiation dose still is an important prognostic factor for LRC.

BACKGROUND: RIPA Buffer exhibits different extraction efficiencies of proteins of cells and tissues, which is not appropriate for all samples. OBJECTIVE: To achieve an optimal lysis buffer for skeletal muscle protein extraction in mice of acidosis, and to provide basis for studies on skeletal muscle atrophy. METHEDS: Twenty male healthy C57BL/6 mice, aged 3 months, weighting 25-30 g, were provided by Laboratory Animal Center of Shanxi Medical University. The mice were sacrificed after anesthesia, and the gastrocnemius muscle of lower extremity was isolated. There were two groups: acidosis group was given 10 g of feed mixed with 0.4 mol/L hydrochloric acid (10 mL) , and control group received 10 g of feed mixed with same volume of water, for 7 consecutive days. The effect of RIPA Buffer, Original Buffer and JP Buffer on the skeletal muscle protein extraction in mice of acidosis was compared. The expression levels of AKT, p-AKT (Thr308) , rpS6 and p-rpS6 (Ser235/236) were detected by western blot assay. GLUT4 mRNA expression was examined by RT-qPCR. RESULTS AND CONCLUSION: (1) Different buffers generated different protein-yields. The protein yield was highest in JP Buffer, but the target protein signal was not high. The protein yield was low in RIPA Buffer. Original Buffer could extract sufficient proteins, and had clear band detected by western blot assay. (2) Western blot assay scores in Original Buffer were higher than those of other two buffers. (3) Western blot assay results showed that the extent of phosphorylation in both groups showed no significant changes. (4) GLUT4 mRNA expression level examined by RT-qPCR showed no significant changes in both groups. (5) These results indicate that Original Buffer is optimal lysate of skeletal muscle protein extraction. Inactivated AKT signaling pathway is seen in the short-term hydrochloric acid-induced acidosis group, so whether lengthening acidosis time can activate the signaling pathway. Selecting the optimal lysis buffer for different samples is premise to ensure western blot assay results.

The integrated curriculums of basic medicine in Shanxi Medical University are as follows: nine basic medical courses were integrated into 10 medical modules, with additional training including PBL and TBL case discussion, clinical clerkship, flipped classroom and other non-integrated subjects etc. In order to assess the interest of students for integrated curriculum, their intrinsic motivation, as well as their comprehension and application of medical knowledge, we gave anonymous questionnaire to 149 students, 16 teachers in basic medicine, and 10 teachers in clinical medicine. Results showed that more than 90% students were willing to take the integrated curriculum and participate in PBL and TBL case discussion, and students who were unwilling to take the curriculum were less than 10%, they thought that the knowledge of new curriculum system was incoherent. The proportion of students from higher grade who were unwilling to participate in the flipped classroom was increased from 6.7% to 95.0%. Most of the teachers both in basic and clinic medicine believed that new curriculum system was helpful for students to comprehend basic medical knowledge and strengthen their discriminating ability, but did not function in improving students' practical ability. In the further reform on teaching, details like the coherence of knowledge and the content selected when self-studying should be improved.

Objective: To investigate the effect of axillary lymph node status in evaluating the prognosis of patients with local recurrence after breast-conserving surgery. Methods: The clinical data of 72 patients with local-regional recurrence who had undergone breast-conserving therapy in Tianjin Cancer Hospital from February 2001 to December 2009 were collected and retrospectively analyzed. These patients were divided into axillary lymph node positive group (23 cases) and negative group (49 cases), according to their axillary lymph node status. Results: Among 72 patients, 21 cases were local recurrence, 35 cases were regional recurrence, and 16 cases were local-regional recurrence. In the axillary lymph node positive group, 7 cases were local recurrence, 10 cases were regional recurrence, 6 cases were local-regional recurrence. In the axillary lymph node negative group, 14 cases were local recurrence, 25 cases were regional recurrence, 10 cases were local-regional recurrence. There was no significant difference between these two groups (P=0.807). Moreover, no significant differences of the age, recurrent site, estrogen receptor (ER) and/or progesterone receptor (PR) status, human epidermal growth factor receptor 2 (HER-2) status, disease-free period, local and systemic therapeutic effect, non-distant metastasis survival between these two groups were observed (P>0.05). However, the overall survival of axillary lymph node positive group after recurrence was significantly lower than that of negative group (P=0.014). Cox multivariate analysis showed that recurrent site is an independent prognostic factor of disease-free survival of patients with regional recurrence after breast-conserving surgery (OR=2.050, P=0.002). The axillary lymph node status and recurrent site were independent prognostic factors of overall survival of these patients (OR=3.469, P=0.003; OR=3.676, P<0.001). Conclusions Axillary lymph node status is an independent factor of overall survival of patients with regional recurrence after breast-conserving surgery, but it is marginally related with their non-distant metastasis survival.