Objective To explore the status of C57BL/6J mouse brown fat adipogenic differentiation function with aging.Methods C57BL/6J female and male mice at the ages of 0-week (newborn),4-week,8-week,12-week old were selected from the same brood,brown adipose tissue was obstained from their interscapular region,and the brown adipose was identified by using immunohistochemical markers.Then the total RNA was extracted from the brown adipose and quality identification was determined at the same time.The expression levels of the related genes (PPARα,C/EBPα,PGC-1α,PPARγ,FOXC2,BMP7) induced by brown adipose adipogenic differentiation were detected by quantitative real-time PCR in 0-week,4-week,8-week,12-week mice.Results Uncoupling protein -1 (UCP1) immunohistochemical data indicated that positive deep-colour substance was brown adipose tissue.Quantitative Real-time PCR also indicated that the expression volume of adipogenesis gene gradually reduced with aging,and there were significant differences at the different time points [PPARα (F =11.96,P ＜ 0.000 1),C/EBPα (F =9.39,P ＜0.000 1),PGC-1α(F =17.21,P ＜0.000 1),PPARγ(F =13.11,P ＜0.000 1),FOXC2(F =12.23,P ＜0.000 1),BMP7(F =16.44,P ＜0.000 1)].Conclusions The adipogenic differentiation ability and activity of mouse brown adipose gradually reduce with aging.But the regulatory factors for its function needs to be further investigated.

Objective To establish an effective DNA isolation method for neonatal disease screening,so as to explore its application to the methylation detection.Methods The 20 dried blood spots samples were randomly divided into 2 groups according to the gender:the traditional method group (n =10) and the improved kit method group(n =10).The DNA quality was evaluated based on its concentration,integrity and whether it could be used in polymerase chain reaction (PCR).These DNA samples with or without bisulfite treatment were used as template in the methylation-specific polymerase chain reaction (MSP).The methylation levels of Leptin and tumor necrosis factor-α (TNF-α) gene promoter region were detected.Results DNA concentration of the improved kit method [(5.70 ± 0.81) mg/L] was significantly higher than that of the traditional method [(3.50 ± 0.45) mg/L] (t =2.79,P ＜ 0.05),and biochemical analyzer analysis showed a better DNA integrity.Agarose gel electrophoresis revealed that 18S gene fragment could be successfully amplified by PCR method,suggesting its potential application to PCR study.MSP results showed different DNA methylation levels of Leptin and TNF-α genes promoter regions from various samples.Conclusions The improved kit method can effectively extract DNA from dried blood spots samples,and these DNA can be used in methylation research.The study can provide a new research direction and technical method to reveal the pathogenesis of disease from the perspective of DNA methylation.

Objective To observe the clinical therapeutic effect of Astragalus injection combined with sub-hibernation therapy for treatment of patients with severe craniocerebral injury.Methods A retrospective research method was conducted, and 89 cases with severe craniocerebral injury admitted to the Department of Critical Care Medicine of the First People Hospital of Baiyin in Gansu Province from January 2010 to December 2016 were enrolled. Astragalus injection combined with sub-hibernation was applied for treatment of 45 patients in the observation group, and for another 44 cases assigned in the control group, simply conventional treatment was used. The Glasgow coma score (GCS), coma time, ratio of cripple, mortality, incidences of pulmonary infection and urinary tract infection were compared between the two groups.Results After treatment, GCS scores were significantly higher than those before treatment in both groups, but the degree of increase was more marked on the 30 days after treatment in the observation group than that in the control group (13.15±2.53 vs. 8.79±1.59,P < 0.05). Compared with the control group, coma time was obviously shorter (days: 5.79±1.89 vs. 13.65±2.73), ratio of cripple [13.33% (6/45) vs. 22.73% (10/44)] and mortality [17.78% (8/45) vs.31.82% (14/44)] were significantly lower in observation group (allP < 0.05); the incidences of pulmonary infection [22.44% (11/45) vs. 31.82% (14/44)] and urinary tract infection [28.89% (13/45) vs. 36.36% (16/44)] were lower in the observation group, but the differences between the two groups were not statistically significant (bothP > 0.05).Conclusions The treatment of Astragalus injection combined with sub-hibernation treatment can significantly improve the coma degree, shorten the time of coma, reduce the mortality and disability rate of patients with severe craniocerebral injury.

Objective To investigate the inlfuence of overexpression of miR-26b on the secretion of adipokines dur-ing human adipocyte differentiation. Methods Human preadipocytes were infected with the hsa-miR-26b over-expressing lentivirus and were induced to differentiate, and then the levels of adipokines (IL-6, leptin, resistin, TNF-α) at different time points during differentiation were measured by ELISA. Results Compared with control group, decreased secretions of both IL-6 and leptin, and increased secretion of resistin were found during the differentiation of human adipocytes in miR-26b overexpressed group. However, the secretion of TNF-αwas not measured in both groups. Conclusion The miR-26b can improve the inlfammation and insulin resistance of human adipocytes, which will provide potential targets for obesity treat-ment.

Objective To observe the effect of Triclosan( TCS) exposure on Caenorhabditis elegans( c. ele-gans) F1 generation of locomotory behavior, brood size, and generation time. Methods The trial included a control group and 4 TCS treatment groups with different doses (100 nmol/L,1 μmol/L,10μmol/L,20μmol/L),the exposure time being 24 hours,the effect of c. elegans′head thrashes,body bending frequency,the brood size and generation time was observed. Results (1) The control group exposed to 100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,their head thrash frequency of c. elegans F1 was(109. 40±8. 61) times/min,(84. 70±7. 82) times/min,(76. 35±7. 44) times/min,(74. 74±5. 93)times/min,(71. 95±4. 19)times/min,respectively,the head thrash ability of c. elegans was significantly inhibited(F=62. 245,P<0. 01). (2) When the control group was exposed to 100 nmol/L,1 μmol/L,10μmol/L,20 μmol/LTCS,the frequency of c.elegans F1 body bent was (19.94±2.46)times/20 s,(15.13±1.99) times/20 s,(14.63±2.31)times/20 s,(14.69±1.96)times/20 s,(12.00±1.86)times/20 s,respectively,and the comparative differences between groups were statistically significant(F=25. 636,P<0. 01). (3) When the control group was exposed to 0,100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,the body sizes of the c. elegans F1 generation was (286.83±6.01)articles,(273.33±6.41)articles,(214.17±7.25)articles,(173.67±9.20)articles, (118. 50 ± 6. 98 ) articles, respectively, the brood size of the C. elegans F1 generation exposed to 100 nmol/L, 1μmol/L,10 μmol/L,20 μmol/L TCS levels,were reduced by 4. 71%,25. 60%,39. 45%,58. 67%,the ge-neration time of the c. elegans′F1 generation was shortened by 2. 14%-5. 38% in the TCS treatment groups compared with the control group(F=27. 520,P<0. 01). Conclusions After c. elegans exposure to TCS,locomotory behavior can be severe-ly affected,reproductive damage causes a decline in the number of brood size,and the speeding-up of the breeding rate is related to the concentration of TCS concentration-response.

Objective To predict the biological process and signaling pathways in which hsa-miR-1908 might be in-volved by a series of bioinformatics analysis, so as to lay foundations and provide theoretical basis for the further studies of hsa-miR-1908 biological function in human preadipocytes. Methods The sequence of hsa-miR-1908 was acquired from miR-Base database, and target genes of hsa-miR-1908 were predicted by miRanda, and then the intersection of the results and the results of gene-chip as gene set were further analyzed by gene ontology and pathway enrichment. Results The hsa-miR-1908 had some conserved property among different species. The functions of the target genes were enriched in Wnt receptor signal-ing pathway through beta-catenin, cell cycle, cell apeptosis and other biological processes. The GnRH signaling, MAPK sig-naling, insulin signaling, cell cycle signal transduction pathways and signaling pathway in pancreatic cancer were signiifcantly enriched. Conclusions The target genes set of hsa-miR-1908 were enriched in multiple biological process which are related with the obesity. This study provides guidance for the further study in human preadipocytes.

Objective To predict the functions of hsa-miR-1908 promoter using various bioinformatic tools, and to provide clues for further study on transcriptional regulation mechanism of miR-1908 in human adipocytes. Methods The promoter se-quence of miR-1908 was obtained from Ensemble, and then the CpG islands and transcription factor binding sites were pre-dicted by a variety of online bioinformatic tools. Results The length of the miR-1908 promoter sequence was 1 458 bp. The CpG islands, which inhibited the transcription of miR-1908, were located at (438-756) bp, (836-937) bp and (979-1374) bp. Meanwhile, 15 transcription factor binding sites were found in the promoter sequence of miR-1908. Conclusions miRNA up-stream promoter related bioinformatics can not only improve the efficiency of microRNA promoter research, but also provide further important information on transcriptional regulation of miR-1908.

Objective To observe the role of Zhenqi Fuzheng granules in the treatment of patients with severe pneumonia respiratory failure. Methods A retrospective research was conducted, 96 patients with severe pneumonia admitted to the Department of Critical Care Medicine of the First People Hospital of Baiyin were enrolled, and they were divided into a western medical conventional treatment group (47 cases) and a combined traditional Chinese and western medicine treatment group (49 cases) according to the different treatment methods. A variety of conventional rescue treatments were carried out in the western medicine routine treatment group (conventional group); while in the combined traditional Chinese and western medicine conventional treatment group (combined group), based on the western medical therapies in the conventional group, at the same time, additionally Zhenqi Fuzheng granules 5 g dissolved in 20 mL warm boiled water were given by nasal feeding to the patients, 2 times daily, 7-10 days constituting a therapeutic course, and after 3 courses, the clinical therapeutic effects were evaluated. The mortality and changes of pre- and post-treatment levels of immunoglobulin, procalcitonin (PCT), C-reactive protein (CRP), course of disease, times of using antibacterial drug and ventilator, and changes in lung exudation before and after treatment, and incidence of pulmonary fibrosis were observed in the two groups. Results The mortality of combined group was significantly lower than that of the conventional group [14.29% (7/49) vs. 36.17% (17/47), P < 0.05]. After treatment, the levels of immunoglobulins were significantly higher than those before treatment, while the levels of PCT and CRP were significantly lower than those before treatment in both groups, and the changes after treatment in the combined group were more obvious than those in conventional group [IgG (g/L): 12.48±2.36 vs. 10.35±3.01, IgA (g/L): 4.16±1.08 vs. 3.18±1.13, IgM (g/L): 5.01±1.15 vs. 3.62±1.02, PCT (mg/L): 1.48±1.36 vs. 2.35±1.01, CRP (mg/L): 8.48±2.36 vs. 10.35±2.01, all P <0.05]. Compared with those in conventional group, the patient's disease course, times of using antibacterial drug and ventilator were significantly shorter in combined group [the course of disease (days): 10.86±2.64 vs. 14.38±3.02, time of using antibacterial drug (days): 11.29±3.54 vs. 13.96±4.21, time of using ventilator (days): 8.26±2.64 vs. 10.25±3.08, all P <0.05]; Compared with those in the conventional group, the cases with significant reduction of pulmonary exudation and the cases with complete absorption of pulmonary exudation were markedly increased, while the cases of pulmonary fibrosis were obviously decreased in the combined group (cases of reduction of pulmonary exudation:15 cases vs. 10 cases, complete absorption of pulmonary exudation: 16 cases vs. 9 cases; cases of pulmonary fibrosis:8 cases vs. 10 cases, all P < 0.05). Conclusion Zhenqi Fuzheng granules can significantly reduce the mortality and inflammatory factor levels, raise immunoglobulin, shorten the course of disease and times of using antimicrobial drugs and ventilator, reduce pulmonary exudation and incidence of pulmonary fibrosis obviously in patients with severe pneumonia.

Objective: To explore the effects of different mild hypothermia therapy time on the efficacy and complications of patients with severe traumatic brain injury (STBI). Methods: A retrospective research method was used. 132 patients with STBI given mild hypothermia therapy admitted to the Third Affiliated Hospital of Gansu University of Traditional Chinese Medicine from January 2010 to December 2018 were enrolled. According to the days of mild hypothermia therapy, the patients were divided into 2-day mild hypothermia treatment group, 5-day mild hypothermia treatment group and 14-day mild hypothermia treatment group. Glasgow coma score (GCS) after treatment of 10 days and 30 days, 30-day disability rate and mortality, coma time, prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), D-dimer, and the incidence of complications such as decreased blood pressure, decreased gastrointestinal motility, infection, nonunion of scalp, cerebrospinal fluid leakage, etc. were compared among three groups. Results: Among the 132 patients with STBI, there were 44 cases in the 2-day mild hypothermia treatment group, 45 cases in 5-day mild hypothermia treatment group, and 43 cases in 14-day mild hypothermia treatment group. There was no significant difference in gender, age, GCS score before treatment or time from injury to admission among three groups. Compared with 2-day mild hypothermia treatment group, the GCS score 10 days and 30 days after treatment in 5-day mild hypothermia treatment group and 14-day mild hypothermia treatment group were significantly higher (11.61±2.23, 10.17±2.03 vs. 6.79±1.49; 13.15±2.53, 11.24±2.24 vs. 8.79±1.59), the coma time were shorten (days: 5.79±1.89, 5.45±1.72 vs. 13.65±2.73), and 30-day disability rate and mortality were significantly decreased [13.33% (6/45), 11.63% (5/43) vs. 22.73% (10/44); 17.78% (8/45), 16.28% (7/43) vs. 31.82% (14/44)], PT and APTT were reduced obviously (s: 20.14±4.12, 22.54±3.56 vs. 30.67±5.19; 35.14±12.41, 38.59±13.54 vs. 56.67±10.62), Fib rose obviously (g/L: 1.84±0.25, 1.98±0.27 vs. 0.67±0.12), and D-dimer reduced obviously (mg/L: 53.10±19.84, 49.20±20.13 vs. 102.60±20.13), with statistically significant differences (all P < 0.05). But there was no significant difference in above indicators between the 5-day mild hypothermia treatment group and 14-day mild hypothermia treatment group (all P > 0.05). The incidence of complications in 14-day mild hypothermia group was significantly higher than those in 2-day mild hypothermia group and 5-day mild hypothermia group [decrease of blood pressure: 55.56% (20/36) vs. 36.67% (11/30), 35.14% (13/37); weakening of stomach intestinemotive power: 72.22% (26/36) vs. 46.67% (14/30), 45.95% (17/37); urethral infection: 52.78% (19/36) vs. 36.67% (11/30), 35.14% (13/37); lungs infection: 47.22% (17/36) vs. 36.67% (11/30), 37.84% (14/37); disunion of scalp: 5.56% (2/36) vs. 0% (0/30), 0% (0/37); leak of cerebrospinal fluid: 5.56% (2/36) vs. 0% (0/30), 0% (0/37), all P < 0.05], but there was no significant difference between the 2-day mild hypothermia treatment group and 5-day mild hypothermia treatment group (all P > 0.05). Conclusions The optimal time frame for mild hypothermia treatment in patients with STBI is 5-day, which shortens the coma time, and reduces the mortality and the disability rate. The shorter mild temperature time cannot effectively prevent secondary brain injury. However, the prolonged period of mild temperature will affect the repair of the patient's injury tissue, which is not conducive to the recovery of patient's mechanical function and is prone to complications.