Objective:To investigate whether 6-shogaol (6-SH) alleviates oxygen-glucose deprivation/reoxygenation (OGD/R)-induced neuronal autophagy and calcium overload by promoting the expression of microRNA-26a-5p (miR-26a-5p) and inhibiting death-associated protein kinase 1 (DAPK1), and to explore its potential mechanisms.Methods:Primary cultured logarithmic growth phase mouse hippocampal neurons HT22 cells were taken and cell counting kit-8 (CCK-8) was used to detect cell viability, searching for the optimal concentration of Na 2S 2O 4. HT22 cells were divided into blank control group (NC group), OGD/R group (sugar-free culture medium + 10 mmol/L Na 2S 2O 4 treatment for 1.5 hours followed by normal culture medium for 4 hours), 6-SH intervention group (cultured with 10 μmol/L 6-SH for 4 hours after OGD), negative control inhibitor pretreatment group (transfected with negative control inhibitor for 48 hours followed by OGD, then cultured with 6-SH for 4 hours), and miR-26a-5p inhibitor pretreatment group (transfected with miR-26a-5p inhibitor for 48 hours followed by OGD, then cultured with 6-SH for 4 hours). Cell viability of each group was detected by CCK-8 method; cell ultrastructure was observed under transmission electron microscopy; real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the gene expressions of DAPK1 and miR-26a-5p; molecular docking were used to verify the interaction between 6-SH and miR-26a-5p; dual-luciferase assay was used to verify the targeting relationship between DAPK1 and miR-26a-5p; flow cytometry was used to determine the levels of intracellular Ca 2+; Western blotting was used to detect the protein expressions of phosphorylated-glutamate receptor 2B (p-NMDAR2B) Ser1303, DAPK1, autophagy related protein Beclin1, light chain 3 (LC3), and p-DAPK1 Ser308; immunofluorescence was used to detect the expression of LC3 and Beclin1. Results:The results of the CCK-8 assay showed that the cell viability of the 6-SH intervention group was significantly increased compared to the OGD/R group, while the cell viability of the miR-26a-5p inhibitor pretreatment group was significantly decreased compared to the 6-SH intervention group. Transmission electron microscopy revealed that the number of autophagosomes in the 6-SH intervention group was significantly reduced compared to the OGD/R group, while the number of autophagosomes in the miR-26a-5p inhibitor pretreatment group was significantly increased compared to the 6-SH intervention group. RT-qPCR results showed that compared with the OGD/R group, the expression of miR-26a-5p was significantly upregulated and the expression of DAPK1 mRNA was significantly downregulated in the 6-SH intervention group; compared with the 6-SH intervention group, the expression of miR-26a-5p was significantly downregulated and the expression of DAPK1 mRNA was significantly upregulated in the miR-26a-5p inhibitor pretreatment group. Molecular docking verified the interaction between 6-SH and miR-26a-5p. Dual-luciferase reporter gene assay showed that compared with the negative control group, mmu-miR-26a-5p significantly downregulated the luciferase expression of m-DAPK1-3UTR-WT, indicating a binding interaction between them. Flow cytometry results showed that compared with the OGD/R group, the level of intracellular Ca 2+ was significantly decreased in the 6-SH intervention group; compared with the 6-SH intervention group, the level of Ca 2+ was significantly increased in the miR-26a-5p inhibitor pretreatment group. Western blotting results showed that compared with the OGD/R group, the protein expressions of p-NMDAR2B Ser1303, DAPK1, Beclin1, and LC3 were significantly decreased in the 6-SH intervention group (p-NMDAR2B Ser1303/β-actin: 2.34±0.27 vs. 4.78±0.39, DAPK1/β-actin: 1.40±0.13 vs. 2.37±0.21, Beclin1/β-actin: 2.61±0.32 vs. 4.32±0.29, LC3/β-actin: 2.52±0.45 vs. 5.09±0.18, all P < 0.05), while the protein expression of p-DAPK1 Ser308 was significantly increased (p-DAPK1 Ser308/β-actin: 0.66±0.09 vs. 0.40±0.02, P < 0.05); compared with the 6-SH intervention group, the protein expressions of p-NMDAR2B Ser1303, DAPK1, Beclin1, and LC3 were significantly increased in the miR-26a-5p inhibitor pretreatment group (p-NMDAR2B Ser1303/β-actin: 4.08±0.14 vs. 2.34±0.27, DAPK1/β-actin: 1.96±0.15 vs. 1.40±0.13, Beclin1/β-actin: 3.92±0.31 vs. 2.61±0.32, LC3/β-actin: 4.33±0.33 vs. 2.52±0.45, all P < 0.05), while the expression of p-DAPK1 Ser308 protein was significantly decreased (p-DAPK1 Ser308/β-actin: 0.33±0.12 vs. 0.66±0.09, P < 0.05); immunofluorescence staining showed that compared with the OGD/R group, the fluorescence intensity of LC3 and Beclin1 was significantly decreased in the 6-SH intervention group; compared with the 6-SH intervention group, the fluorescence intensity of LC3 and Beclin1 was significantly increased in the miR-26a-5p inhibitor pretreatment group. Conclusion:6-SH can alleviate neuronal damage by regulating miR-26a-5p/DAPK1 to reduce autophagy and calcium overload in cells.

Objective: To investigate the mechanism of electroacupuncture (EA) for treating depression and to explore the role of brevican in the medial prefrontal cortex (mPFC) in modulating stress susceptibility and the antidepressant effects of EA in rats. Methods: Twenty-four Sprague–Dawley (SD) rats were equally divided into three groups: green fluorescent protein (GFP) + control, GFP + chronic unpredicted mild stress (CUMS), and short-hairpin RNA targeting on brevican (shBcan) + CUMS. Another 24 SD rats were equally divided into CUMS + GFP, CUMS + GFP + EA, and CUMS + shBcan + EA groups. Behavioral tests were conducted to assess depression-like behavior. Western blot analysis was used to evaluate the expression of brevican, aggrecan, GLuA1, and PSD95 in mPFC subregions. Results: Behavioral parameter evaluation show that rats in the shBcan + CUMS group exhibited a significantly reduced sucrose preference (P = .0002) and increased immobility time (P = .0011) compared to those in rats in the GFP + CUMS group. Western blotting showed that brevican expression was significantly downregulated in the PrL of the shBcan + CUMS group compared with that in the GFP + CUMS group (P = .0192). Furthermore, compared to the CUMS + GFP + EA group, the CUMS + shBcan + EA group exhibited a significantly decreased sucrose preference (P = .0334), increased immobility time (P = .0465), and increased latency to food (P = .0261). In the CUMS + shBcan + EA group, the EA-induced brevican and PSD95 overexpression was reversed, compared with that in the CUMS + GFP + EA group (P = .0454 and P = .0198, respectively). Conclusion EA exerts its antidepressant effects through the modulation of brevican expression in rats. Our findings highlight the important role for brevican in stress susceptibility, which could be a potential target for treating depression.

Objective:To construct a targeted and accurate evaluation system for facial and cervical wounds and scars of burn patients.Methods:The method combining literature analysis and survey research was adopted, and the basic principles of item system construction were followed. From June to August 2020, based on the aesthetic standards of facial and cervical plastic surgery, the topographic map assessment system for facial and cervical wounds and scars of burn patients was preliminarily formed, focusing on the assessment of wounds and scars in the necks and faces of patients after burns. In September 2020, 38 experts in the relevant fields were consulted in advance and the questionnaire was revised according to the experts' opinions. From December 2020 to March 2021, the Delphi method was applied to conduct inquiry by correspondence with 35 experts in relevant fields from Guangzhou, Shenzhen, Shanghai, Beijing, and other cities, who met the inclusion criteria, and the items were screened and established. The effective recovery rate of inquiry questionnaire was calculated to determine the level of enthusiasm of experts, the average authority coefficient of all items was calculated to determine the level of expert authority, the average importance expert score, the average coefficient of variation, and the average full score rate of all the third-level items were calculated to determine the concentration of expert opinions, the average coefficients of variation and Kendall's harmony coefficients of the importance, sensitivity, and operability expert scores of all the third-level items were calculated to determine the degree of coordination of expert opinions. The Kendall's harmony coefficients for the importance, sensitivity, and operability expert scores of all the third-level items were statistically analyzed with chi-square test.Results:Among the 35 experts consulted by Delphi method, mainly were male, aged (48±10) years, with 8-38 years of working experience, mainly with associate senior titles and above, all with a bachelor's degree or above education background, and of whom 11 were burn experts, 7 were wound repair experts, 4 were plastic surgery experts, and 13 were rehabilitation medicine experts. Finally, a topographic map assessment system for facial and cervical wounds and scars of burn patients was formed, including 4 first-level items, 21 second-level items, 40 third-level items, and 1 mask. The effective recovery rate of inquiry questionnaire was 100% (35/35). The average authority coefficient of all items was 0.89. The average importance expert score was 4.67, the average coefficient of variation of importance expert score was 0.01, and the average full score rate of all the third-level items was 86.3%. The average coefficients of variation of the importance, sensitivity, and operability expert scores of all the third-level items were 0.01, 0.01, and 0.02, respectively. The Kendall's harmony coefficients for the importance, sensitivity, and operability expert scores of all the third-level items were statistically significant (with χ2 values of 1 201.53, 745.67, and 707.07, respectively , P<0.05). Conclusions:The established topographic map assessment system for facial and cervical wounds and scars of burn patients has high scientificity and reliability, which can be used for the evaluation of facial and neck wounds or scars in burn patients.

Objective: To evaluate the safety of a herbal pilatory for external use. Methods : An acute eye irritation test were employed to detect the eye irritation of the herbal pilatory;a skin irritation test,a skin sensitization test and a skin phototoxicity test were employed to detect the dermal toxicity;Salmonella typhimurium reverse mutation assay（Ames test）and chromosome aberration test in CHL cells were employed to detect the effects of the pilatory on gene mutation and chromosome aberration in prokaryotic and eukaryotic cells. Results: When the eyes of rabbits exposed to the pilatory without rinse during the first 24 hours,the conjunctiva showed congestion and edema with the highest score of 2,corneal opacity was observed with the highest score of 1;however,these symptoms returned to normal within 72 hours,with the score reduced to 0. No irritation to the skin of rabits was found after exposed to the pilatory for fourteen days,no skin sensitization was introduced by Buehler test and no skin phototoxicity on guinea pigs was detected. There was no abnormal growth of reverse mutation colonies induced by the pilatory under S9 acitivation or not. There was no statistically significant rise of chromosome aberration rate in the exposed CHL cells compared to the control groups（P>0.05）. Conclusion Under the condition,the herbal pilatory showed mild and reversible irritation to eyes,while no dermal toxicity and genetic toxicity were observed. The safety of the herbal pilatory for external use is acceptable.

We prepared a polymorphic form of valnemulin hydrogen tartrate (Form I) to overcome the instability and irritating odor of valnemulin hydrochloride that affect its use in the production and application of veterinary drugs. The physicochemical properties of Form I were characterized by scanning electron microscopy, X-ray powder diffraction, infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. The results showed the crystal structure and thermal properties of Form I were very different from those of a commercially available form of valnemulin hydrogen tartrate (Form II). Form I and Form II were more stable than valnemulin hydrochloride after storage under irradiation and high humidity conditions, respectively. The solubility of Form I was 2.6 times that of Form II, and Form I was selected for use in pharmaceutical kinetics experiments in vivo. Compared to valnemulin hydrochloride, after oral administration at a dose of 10 mg/kg in pigs, Form I had similar pharmaceutical kinetic behavior but a slightly higher area under the concentration–time curve from time zero to the last measurable concentration. Consequently, Form I should be suitable for the development of simple formulations and be effective in the clinical application of veterinary drugs.
Administration, Oral ; Calorimetry, Differential Scanning ; Humidity ; Hydrogen ; Kinetics ; Microscopy, Electron, Scanning ; Odors ; Pharmacokinetics ; Powder Diffraction ; Solubility ; Spectrum Analysis ; Swine ; Veterinary Drugs

Objective To validate a chemical shift-encoded MRI(CSE-MRI)water-fat imaging for quantifying vertebral marrow fat content using MRS as the reference standard.Methods MRS and CSE-MRI were performed to calculate proton density fat fraction(PDFF) in 83 subjects,including 41 normal bone mass,26 osteopenia and 16 osteoporosis.Eight participants were scanned three times with repositioning to assess the repeatability of CSE-MRI PDFF measurements.Agreements of intra-observer and inter-observer were evaluated by intraclass correlation coefficient(ICC).Linear regression,Bland-Altman 95% limit of agreement and Lin's concordance correlation coefficient were calculated.Results The repeatability for CSE-MRI PDFF measurements expressed as absolute precision error was 1.45%.PDFF was 62.1%±11.1% by MRS and 60.4%±10.1% by CSE-MRI in 83 subjects.There were significant differences in PDFF among the normal bone mass,osteopenia and osteoporosis groups after adjusting for age,years since menopause and body mass index (all P<0.001).The intra-and inter-rater reliability for duplicate measurements at CSE-MRI PDFF were more than 0.993.Pearson correlation coefficient was 0.979 and Lin's concordance correlation coefficient was 0.962.All data points calculated using the Bland-Altman method were within the limits of agreement.Inverse associations were observed between BMD (r=-0.560--0.710)and CSE-MRI-based PDFF,and between BMD (r=-0.539--0.706)and MRS-based PDFF in various groups.Conclusion CSE-MRI with multiple lipids peak model and T2?-correction is equally accurate in characterizing marrow fat content as MRS.

In this study,the intestinal microbial flora diversity of adult and young African lions in the same breeding environment was detected by PCR-DGGE technique.Total bacterial DNA was extracted and 16S rDNA V3 region was amplified,then conducting PCR-DGGE.Subsequently,the specific bands of DGGE were cloned and sequenced.The bacterial species were identified by comparing the sequence through BLAST.The results indicated that the intestinal microbial flora of adult African lions includes Clostridium,Lachnospiraceae bacterium,Anaerovorax,Lactococcus,Peptostreptococcus and Blautia.While the intestinal microbial flora of young African lions is lesser,most bacteria are common to adult and young lions,such as Bacteroidetes bacterium and rumen bacterium.The UPGMA clustering analysis of the DGGE fingerprint showed the similarities of the bacteria structures between adult and young African lions were only 34％.These results revealed that the intestinal microbial flora has significant difference in different stages of African lions.This study lays a foundation for the development of microecological agents in different growth stages of wild animals.

Objective To investigate the interest,cognition degree,participation degree and current situation of the advanced undergraduate clinical students in our college,to discuss the reason for low level of students' scientific research quality,in order to provide the reference to making research training programs for them.Methods The advanced undergraduates of seven-year and five-year clinical program interned in orthopedics department from January to June in 2014 were selected with cluster sampling method,and investigated anonymously by questionnaires and interview.120 students were investigated by questionnaire,and 120 effective questionnaires were taken back.30 students were interviewed.Results The results showed that 90.0 percent (n=108) of students were interested in scientific activity,and 47.5 percent (n=57) of students had participated in scientific lectures.In interviews,students think factors hindering the research on the participation in scientific research are:1)too many courses and heavy school tasks;2) absence of relative knowledge;3) lack of support from college;4) tough condition for scientific training;5) immature management system.Conclusion The advanced undergraduate clinical students had great interest in scientific activity,but had few opportunities to take part in,leading to their low level of scientific research quality.A variety of measures should be taken to bolster their scientific training.

OBJECTIVETo investigate atmospheric mercury concentration in the workplace and urinary mercury concentration in workers exposed to mercury in a thermometer factory, and to determine the levels and influencing factors of urinary Β₂-microglobulin (Β₂-MG) and retinol-binding protein (RBP) in these workers.

METHODSAn occupational health survey of the workplace was completed according to relevant national occupational health standards. Questionnaire survey and occupational health examination were conducted in 178 workers exposed to mercury in the factory. Statistical analysis was accomplished using SPSS 19.0.

RESULTSIn the workplace, atmospheric mercury concentration was out of limits at seven of eight detection points expressed by short-term exposure limit; it was out of limits at all the eight detection points shown by time-weighted average. Statistically significant difference in atmospheric mercury concentration was found among different detection points (F = 138.714, P < 0.001). The geometric mean of urinary mercury concentration measured in 154 workers was 171.607 µg/g. There were 127 workers with urinary mercury concentration exceeding the standard (82.5% over-standard rate). Significant difference in urinary mercury concentration was shown in the workers among different positions (χ² = 44.531, P < 0.01). Urinary mercury concentration was positively correlated with atmospheric mercury concentration (r = 0.624, P < 0.01). The mean urinary Β₂-MG level measured in 148 workers was 0.142 mg/L, and seven workers had urinary Β₂-MG levels greater than 0.3 mg/L (4.7% abnormal rate). The mean urinary RBP level measured in 153 workers was 0.485 mg/L, and 19 workers had urinary RBP levels greater than 0.7 mg/L (12.4% abnormal rate). Ordinal logistic regression showed that age >34 years (OR = 4.88, 95%CI: 2.24∼10.62) and length of service >15 years (OR = 2.50, 95%CI: 1.06-5.92) were risk factors for increased urinary Β₂-MG level. Age >45 years (OR = 7.52, 95%CI: 2.50∼22.65) was a risk factor for increased urinary RBP level.

CONCLUSIONIn the thermometer factory under study, atmospheric and urinary mercury concentrations both seriously exceeded the standards, which were harmful to the health of workers. High atmospheric mercury concentration, old age, and long length of service were risk factors for increased urinary Β₂-MG and RBP levels in workers exposed to mercury.

Adult ; Environmental Exposure ; Humans ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; Mercury ; analysis ; toxicity ; Occupational Exposure ; Risk Factors ; Threshold Limit Values ; Time Factors ; Workplace

Objective To evaluate the effect of postural drainage assisting trachea suction on meconium aspiration syndrome. Methods Total 61 cases of asphyxia neonates with MAS who were born in our hospital from Jan,2007 to Dec,2008, were divided into control group (24 cases) and observing group( 37 cases). The neonates in control group had endotracheal suction directly after intubation. But the infant in observing group was treated with endotracheal suction after postural drainage. The amount of suction from endotracheal tube,the complication of MAS and the outcome of these newborns were evaluated. Results The total amount of meconium drainage from endotrachea in observing group was statistics significantly more than that in control group [( 2. 16 ± 1.82) ml vs ( 1.23 ± 0. 97 ) ml, P ＜ 0. 05 )]; The intubating times in observing group were statistical significantly less than that in control group[( 1.19 ± 0. 46) vs ( 1.79 ± 0. 83 ) times, P ＜0. 01 ). The incidence of complication in observing group was 8. 11% ,which was significantly lower than that in control group(29. 17% ,P ＜0. 05). There were shorter needing oxygen time [(21.30 ± 22. 38) h vs (52. 91 ±39. 20) h,P ＜0. 01]and shorter hospitalization days [(9. 24 ±3.94) d vs ( 14. 39 ±6. 49) d,P ＜0.01 )]in observing group than those in control group respectively. The mortality in control group was 4. 17%, and no death occurred in observing group. Apgar scores of the first minute was similar in both groups ( P ＞ 0. 05 ). But there was significant difference(70. 16% vs 58. 34% ,respectively;P ＜0. 05) in the fifth minute Apgar scoring of 8 ～ 10 scores between the observing group and the control group. Conclusion Postural drainage assisting endotracheal suction may remove meconium in trachea effectively, decrease the complications of MAS and shorten the oxygen days and hospitalization time.