Objective To construct and identify replication deficient recombinant adenovirus expressing human c-Jun N-terminal kinase(JNK)by homologous recombination adenovirus dominant-negative type JNK(Ad-DN-JNK).Methods The linearized recombinant shuttle vector pAdTrack-CMV-DN-JNK Was co-transformed with backbone vector pAdEasy-l into bacteria BJ5183 for recombinant adenoviral vector.The recombinant adenoviral vector was transfected into HEK293 packing cells tO construct replication deficient recombinant adenovirus,and then the recombinant edenovirns WaS detected by PCR and DNA sequencing.Western blot analysis was utilized to detect the Cxpression of Ad-DN-JNK and the level of insulin receptor substrate l Serine307 phosphorylation.Results JNK recombinant adenoviral vectorcould be effectively transfeeted into HEK 293 cell and successfully packed by intracellular enzyme.The expression of green fluorescent protein(GFP)Was observed on the 5th day after transfection.The fragment of JNK gene waS amplified by PCR and identified by sequencing.The titer of the prepared Ad-DN-JNK is 2.5×1010 pfu/ml.The animal experiment confirmed that constructed Ad-DN-JNK could be effectively expressed in liver tissue.Conclusion The research successfully constructed recombinant adenoviral vector and recombinant adenoviral particle.Animal experiment demonstrated the Ad-DN-JNK could effectively mediated the expression of DN-JNK gene and down-regulated the level of IRSlscfine307 phosphorylation.The achievement laid a foundation for further investigation of the function and application of JNK.

Objective To construct replication deficient recombinant adenovirus expressing human c-Jun N-terminal kinase by homologous recombination.Methods The linearized recombinant shuttle vector pAdTrack-CMV-WT-JNK was co-transformed with backbone vector pAdEasy-1 into bacteria BJ5183 for recombinant adenoviral vector.The recombinant adenoviral vector was transfected into HEK293 packing cells to construct replication deficient recombinant adenovirus,and then the recombinant adenovirus was detected by PCR and DNA sequencing.Results JNK recombinant adenoviral vector was effectively transfected into HEK 293 cells and was successfully packed by intracellular enzyme.The expression of green fluorescent protein(GFP)was observed on the 5th day after transfection.The fragment of JNK gene was amplified by PCR and identified by sequencing.The animal experiment confirmed that Ad-WT-JNK was effectivety expressed in liver tissue. Conclusion The research successfully constructed recombinant adenoviral vector and recombinant adenoviral particle.And the achievement laid a foundation for further investigation of the function and application of JNK.

Objective To explore the effect of damage control surgery (DCS) in the treatment of severe electric burn. Methods Retrospective analysis on clinical data of 45 patients with severe electric burn was con-ducted. According to implementing DCS or not , patients were separated into DCS group and control group. In DCS group, tangential excision and transplanted xenogenic acellular dermal matrix was conducted for severe electric burn cases with deep Ⅱ degree wound, and escharectomy and VSD dressing for Ⅲ~Ⅳ degree electric contact burn wound at the first stage then skin-grafting or skin flap-grafting on the secong stage was applied. For control group , debridement, tangential excision or escharectomy and skin-grafting or skin flap-grafting to close the wound were conducted. We compared the difference in terms of operation time, length of stay, disability rate, mortality and complications between 2 groups. Results The operation time, incidince of disability and complications in DCS Group obviously decreased but there was no difference in length of stay and mortality in both groups. Conclusion DCS is effective for reducing complications and optimizing therapeutic effect for severe electric burn patients.

Objective To explore mediating effect of coping style on the relationship between self-esteem and negative emotions among college students with left-behind experience. Methods The investiga-tion was carried out among 663 college students with left-behind experience by multistage sampling method, using coping style scale, self-esteem scale, self-rating anxiety scale and Beck depression inventory. Re-sults Scores of all scales were shown as follows: self-esteem (3. 09±0. 51),anxiety (34. 94±7. 90),de-pression (8. 80±8. 72),solving problem coping (9. 03±2. 57),help-seeking coping (5. 70±2. 24),self-bla-ming coping (3. 54±2. 24),fantasy coping (4. 56±2. 35),avoidance coping (4. 74±2. 02),rationalizing coping (4. 67±2. 32). Anxiety and depression had significantly negative correlations with self-esteem(r=-0. 46,-0. 50,P<0. 01) ,solving problem coping( r=-0. 29,-0. 30,P<0. 01) and help-seeking coping( r=-0. 08,-0. 18,P<0. 01) ,and significantly positive correlations with rationalizing coping( r=0. 32,0. 28,P<0. 01),self-blaming coping(r=0. 45,0. 43,P<0. 01),fantasy coping(r=0. 33,0. 23,P<0. 01) and avoid-ance coping(r=0. 26,0. 23,P<0. 01). There were partially significant mediating effects of solving problem coping(-0. 09,95％CI:-0. 06--0. 13),rationalizing coping(-0. 03,95％CI:-0. 01--0. 05),self-blaming coping(-0. 11,95％CI:-0. 06--0. 15) ,fantasy coping(-0. 02,95％CI:-0. 01--0. 04) ,and non-significant effects of help-seeking coping and avoidance coping in the relationship between self-esteem and negative e-motions. Conclusion Solving problem coping,rationalizing coping,self-blaming coping and fantasy coping play a partial mediating role in the relationship between self-esteem and negative emotions,while the media-ting effect of help-seeking coping and avoidance coping are not significant.

OBJECTIVE:To provide reference for further promoting the clinical application of chronopharmacology and rational drug use.METHODS:The self-designed questionnairea was used to conduct on-site investigation among participants from 40 hospitals in Anhui province by dint of a meeting.The survey data were analyzed statistically.RESULTS:Totally 170 questionnaires were distributed,and 166 were retrieved,the total response rate was 97.65％.93.37％ of the interviewed pharmacists were aware of the circadian rhythms,and 87.35％ were aware of the circadian rhythms of more than two kinds;interviewed pharmacists expressed better awareness of indicators like gastric acid secretion (75.30％),blood pressure (67.47％) and growth hormone (66.87％) etc.while their awareness of other diseases like anaphylactic rhinitis (66.87％),gastric ulcer (50.60％) and migraine (50.00％) remained to improve.All the interviewed pharmacists recognized the better therapeutic efficacy (84.34％) and lower side effect (81.33％) of chronopharmacology-referred medication;chronopharmacology characteristics of glucocorticoid drugs were best known (34.34％),followed by antihypertensive drugs (25.30％).CONCLUSIONS:Pharmacists in Anhui province have certain knowledge of chronopharmacology such as biological rhythms,biological process or indicators,biological rhythms of disease and advantage of chronopharmacology-referred medication.It indicates that there has been a foundation for popularizing the concept of chronopharmacology in clinical medication in China;but not often contacted physiological processes or indexes,the understanding of biological rhythm of uncommon diseases are still insufficient,and the understanding of chronopharmacology characteristics of various drugs are far from ideal.

Objective:To establish a multi-factor prediction model for vascular aging based on psychological behavior and physical exercise using nomogram and structural equation method.Methods:A total of 701 inpatients or physical examination patients who underwent carotid ultrasound examination in the Affiliated Hospital of Qingdao University from October 2021 to March 2023 were selected by cross-sectional method. The subjects were randomly divided into training queue( n=492) and verification queue( n=209) according to the ratio of 7∶3.The psychological and behavioral factors, lifestyle, demography and accompanying diseases data were collected. Multi-factor binary Logistic regression method in SPSS 25.0 software was used to screen independent influencing factors of vascular aging, R 4.2.2 software was used to build a nomogram prediction model of vascular aging, and the ROC curve, calibration chart and decision curve analysis were used to evaluate the nomogram, and AMOS software was used to build a structural equation model of vascular aging. Results:Multivariate Logistic regression analysis showed that type A behavior( B=1.757, OR(95% CI)=5.790(2.750-12.210), P<0.001), physical exercise( B=-3.019, OR(95% CI)=0.050(0.020-0.100), P<0.001), high-fat diet( B=0.679, OR(95% CI)=1.970(1.350-2.880), P<0.001), sleep quality( B=-1.451, OR(95% CI)=0.230(0.120-0.460), P<0.001), tea drinking habits( B=-2.349, OR(95% CI)=0.100(0.050-0.200), P<0.001), age( B=0.061, OR(95% CI)=1.060(1.020-1.110), P=0.005), sex( B=-1.263, OR(95% CI)=0.280(0.140-0.570), P<0.001), hyperlipidemia( B=0.679, OR(95% CI)=1.970(1.350-2.880), P<0.001) and diabetes( B=0.838, OR(95% CI)=2.310(1.140-4.700), P=0.021)were independent influencing factors of vascular aging. The nomogram model showed that type A behavior, physical exercise, high-fat diet, sleep quality, tea drinking habits, age, sex, hyperlipidemia and diabetes were independent influencing factors of vascular aging.Advanced age, type A behavior, hyperlipidemia, diabetes and the score of high-fat diet were risk factors for vascular aging, while better sleep quality, tea drinking habit and the score of physical exercise were protective factors for vascular aging.Men were more prone to vascular aging than women. Structural equation model showed that type A behavior had the strongest direct positive effect on vascular aging (the effect coefficient=0.197, P<0.01), and physical exercise had the strongest direct negative effect on vascular aging (the effect coefficient=-0.452, P<0.01), which could indirectly affect vascular aging through various factors. Conclusions:The nomogram model shows the independent influencing factors of vascular aging and has certain predictive value.Structural equation model shows that many factors such as psychological behavior and lifestyle can directly or indirectly affect the occurrence of vascular aging, among which type A behavior and physical exercise have more extensive effects.

Circadian clocks are the internal time-keeping mechanisms for organisms to synchronize their cellular and physiological processes to the daily light/dark cycles. The molecular mechanisms underlying circadian clocks are remarkably similar in eukaryotes. Neurospora crassa, a filamentous fungus, is one of the best understood model organisms for circadian research. In recent years, accumulating data have revealed complex regulation in the Neurospora circadian clock at transcriptional, posttranscriptional, post-translational and epigenetic levels. Here we review the recent progress towards our understanding of the molecular mechanism of the Neurospora circadian oscillator. These advances have provided novel insights and furthered our understanding of the mechanism of eukaryotic circadian clocks.
Circadian Clocks ; Epigenomics ; Neurospora ; genetics ; metabolism ; physiology ; Neurospora crassa ; genetics ; metabolism ; physiology

Abstract OBJECTIVE To establish an efficient and simple HPLC-MS/MS method for determination of flucloxacillin in newborn plasma, and to investigate the interaction between ambroxol and flucloxacillin in newborns. METHODS The samples were analyzed by API4000 HPLC-MS/MS. Ultimate XB-C18 column(2.1 mm×100 mm, 5 μm) were carried out. The mobile phase was composed of water-0.1% formic acid(A) and acetonitrile-0.1% formic acid(B). The quantitative analysis of the ion transitions were monitored at m/z 452.6→284.2 for flucloxacillin and m/z 821.4→397.3 for rifampicin(internal standard). RESULTS The linear range of flucloxacillin under this analysis method was 0.20-80 ng·mL-1, and the lower limit of quantification was 0.20 ng·mL-1; The intra-day and inter-day precision of flucloxacillin were both less than 8.23%; The extraction recovery was in the range of 85.3%-89.2%, and the matrix effect was between 89.3%-92.3%; The stability of plasma samples was good under conditions of 12 h at room temperature, 4 h at room temperature after treatment, repeated freeze-thaw for 3 times, and -20 ℃ freezing for 30 d. The results of clinical samples indicated that the combination of ambroxol could significantly increase the blood concentration of flucloxacillin. CONCLUSION The established HPLC-MS/MS method is accurate, sensitive and can be used for the determination of flucloxacillin concentration in neonatal plasma. The results of clinical samples indicate that ambroxol can significantly increase the blood concentration of flucloxacillin. There are drug interactions between ambroxol and flucloxacillin.

ObjectiveTo study the protective effect of lipoic acid (LA) on the spermatogenic function of the male rats with oligoasthenozoospermia induced by ornidazole (ORN).

METHODSSeventy male SD rats were equally randomized into groups A (solvent control: 1 ml 0.5% CMC-Na + 1 ml olive oil), B (low-dose ORN model: 400 mg/kg ORN suspension + 1 ml olive oil), C (low-dose ORN + low-dose LA treatment: 400 mg/kg ORN + 50 mg/kg LA), D (low-dose ORN + high-dose LA treatment: 400 mg/kg ORN + 100 mg/kg LA), E (high-dose ORN model: 800 mg/kg ORN suspension + 1 ml olive oil), F (high-dose ORN + low-dose LA treatment: 800 mg/kg ORN + 50 mg/kg LA), and G (high-dose ORN + high-dose LA treatment: 800 mg/kg ORN + 100 mg/kg LA), and treated respectively for 20 successive days. Then all the rats were sacrificed and the weights of the body, testis, epididymis and seminal vesicle obtained, followed by calculation of the organ index, determination of epididymal sperm concentration and motility, and observation of the histomorphological changes in the testis and epididymis by HE staining.

RESULTSCompared with group A, group E showed significantly decreased body weight (［117.67 ± 11.53］ vs ［88.11 ± 12.65］ g, P < 0.01) and indexes of the testis (［1.06 ± 0.12］ vs ［0.65 ± 0.13］ %, P < 0.01) and epididymis (［0.21 ± 0.03］ vs ［0.17 ± 0.01］ %, P < 0.01). In comparison with group E, group F exhibited remarkable increases in the epididymal index (［0.17 ± 0.01］ vs ［0.20 ± 0.02］ %, P < 0.01), and so did group G in the body weight (［88.11 ± 12.65］ vs ［102.70 ± 16.10］ g, P < 0.05) and the indexes of the testis (［0.65 ± 0.13］ vs ［0.95 ± 0.06］ %, P < 0.01) and epididymis (［0.17 ± 0.01］ vs ［0.19 ± 0.02］ %, P < 0.05), but no obvious difference was observed in the index of seminal vesicle among different groups. Compared with group A, group B manifested significant decreases in sperm motility (［74.12 ± 8.73］ vs ［40.25 ± 6.08］ %, P < 0.01), and so did group E in sperm count (［38.59 ± 6.40］ vs ［18.67 ± 4.59］ ×105/100 mg, P < 0.01) and sperm motility (［74.12 ± 8.73］ vs ［27.58 ± 8.43］ %, P < 0.01). Sperm motility was significantly lower in group B than in C and D (［40.25 ± 6.08］ vs ［58.13 ± 7.62］ and ［76.04 ± 8.44］%, P < 0.01), and so were sperm count and motility in group E than in F and G (［18.67 ± 4.59］ vs ［25.63 ± 9.66］ and ［29.92 ± 4.15］ ×105/100 mg, P < 0.05 and P < 0.01; ［27.58 ± 8.43］ vs ［36.56 ± 11.08］ and ［45.05 ± 9.59］ %, P < 0.05 and P < 0.01). There were no obvious changes in the histomorphology of the testis and epididymis in groups A, B, C and D. Compared with group A, group E showed necrotic and exfoliated spermatogenic cells with unclear layers and disorderly arrangement in the seminiferous tubules and remarkably reduced sperm count with lots of noncellular components in the epididymal cavity, while groups F and G exhibited increased sperm count in the seminiferous tubules and epididymis lumen, also with exfoliation, unclear layers and disorderly arrangement of spermatogenic cells, but significantly better than in group E.

CONCLUSIONSLA can reduce ORN-induced damage to the spermatogenetic function of rats, improve sperm quality, and protect the reproductive system.

Animals ; Antioxidants ; pharmacology ; Asthenozoospermia ; chemically induced ; drug therapy ; Body Weight ; drug effects ; Epididymis ; anatomy & histology ; drug effects ; Male ; Oligospermia ; chemically induced ; drug therapy ; Ornidazole ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminal Vesicles ; anatomy & histology ; drug effects ; Seminiferous Tubules ; anatomy & histology ; drug effects ; Sperm Count ; Sperm Motility ; drug effects ; Spermatogenesis ; drug effects ; Spermatozoa ; drug effects ; Testis ; anatomy & histology ; drug effects ; Thioctic Acid ; pharmacology