Objective To investigate the effect and mechanism of action in pancreatic cancer BxPC3 cell treated by cerulenin in combination with gemcitabine.Methods BxPC3 cells were cultivated with 10 μg/ml cerulenin,20 μmol/L gemcitabine or 10 μg/ml cerulenin + 20 μmol/L gemcitabine for 48 h,and cells without treatment were control.Cell proliferation was detected by CCK-8 assay,and early apoptosis was detected by AnnexinV/PI double staining method.The expression of Bcl-2 mRNA and Bax mRNA were detected by RT-PCR,and the protein level of Bcl-2,Bax were detected by Western Blot.Results The inhibition rate of BxPC3 cells were 0,(51.28 ± 1.84) ％,(53.59 ± 1.62) ％,(84.57 ± 1.01) ％ in control,cerulenin group,gemcitabine group,combination group; and the rate of early apoptosis was (0.83 ± 0.31) ％,(31.37 ± 1.04) ％,(38.33 ± 0.75) ％,(69.43 ± 0.83) ％,and the expression of Bcl-2 mRNA was 0.67 ± 0.01,0.44 ±0.01,0.36 ±0.08,0.27 ±0.07,and the expression of Bax mRNA was 0.14 ±0.01,0.31 ± 0.02,0.32 ± 0.03,0.91 ± 0.06 ; while the ratio of Bcl-2 mRNA/Bax mRNA was 4.78 ± 0.13,1.39 ± 0.04,1.15 ± 0.02,0.30 ± 0.02 ; the expression of Bcl-2 protein was 1.24 ± 0.04,0.51 ± 0.02,0.42 ± 0.02,0.13 ±0.01 ; and the expression of Bax protein was 0.20 ± 0.05,0.47 ± 0.01,0.54 ± 0.01,1.21 ± 0.03 ; while the ratio of Bcl-2/Bax was 6.00 ± 0.11,1.11 ± 0.01,0.77 ± 0.03,0.10 ± 0.06.And the difference among the groups was statistically significant (P ＜ 0.01).Conclusions Cerulenin combined with gemcitabin has synergistic effect on the inhibition of BxPC cells proliferation.Its mechanisms may be up-regulation of Bax,down-regulation of Bcl-2,and promoting apoptosis of cells through the decrease of Bcl-2/Bax ratio.

The hybrid pencil beam model (HPBM) is an effective algorithm for calculating electron dose distribution in radiotherapy. The mean energy distribution of incident electron beam in phantom is one of the factors that affect the calculation accuracy of HPBM, especially in field edge areas near the end of the electron range. A new fitted formula based on Monte Carlo (MC) simulation data for electron beams with energy range of 6-20 MeV in the homogeneous water phantom is proposed in this paper. The precision of the fitted formula within the scope of the energy was evaluated by comparing the electron dose distribution of ECWG measured data with that obtained from HPBM which took the mean electron energy that calculated by the fitted formula and the existed empirical formula, respectively. The results showed that the accuracy of dose distribution that obtained by the mean electron energy calculated with the fitted formula increased about 1%.
Algorithms ; Electrons ; Humans ; Monte Carlo Method ; Phantoms, Imaging ; Radiotherapy Planning, Computer-Assisted ; Water

We constructed plasmid pMTac to overexpress 3-ketosteroid-Δ1-dehydrogenase (KSDD) in Mycobacterium neoaurum JC-12 for improving androst-1,4-diene-3,17-dione (ADD) production. To construct pMTac, pACE promoter on pMF41 was replaced by tac promoter, and then four recombinants were constructed, which were M. neoaurum JC-12/pMF41-gfp, M. neoaurum JC-12/pMTac-gfp, M. neoaurum JC-12/pMF41-ksdd and M. neoaurum JC-12/pMTac-ksdd. Fluorescence detection results show that much more green fluorescent protein (GFP) was expressed in M. neoaurum JC-12/pMTac-ksdd than M. neoaurum JC-12/pMF41-ksdd. The activity of KSDD was 2.41 U/mg in M. neoaurum JC-12/pMTac-ksdd, 6.53-fold as that of M. neoaurum JC-12 and 4.36-fold as that of M. neoaurum JC-12/pMF41-ksdd. In shake flask fermentation, ADD production of M. neoaurum JC-12/pMTac-ksdd was 5.94 g/L, increased about 22.2% compared to the original strain M. neoaurum JC-12 and 12.7% to M. neoaurum JC-12/pMF41-ksdd. AD (4-androstene-3,17-dione) production of JC-12/pMTac-ksdd was 0.17 g/L, decreased 81.5% compared to M. neoaurum JC-12 and 71.2% to M neoaurum JC-12/pMF41-ksdd. In the 5 L fermenter, 20 g/L phytosterols was used as substrate, ADD production of M. neoaurum JC-12/pMTac-ksdd was improved to 10.28 g/L. pMTac is favorable for expressing KSDD in M. neoaurum JC-12, and overexpression of KSDD has beneficial effect on ADD producing, and it is the highest level ever reported using fermentation method in M. neoaurum.
Androstadienes ; metabolism ; Fermentation ; Industrial Microbiology ; Mycobacterium ; Oxidoreductases ; genetics ; metabolism ; Phytosterols ; metabolism ; Plasmids

The single molecule imaging and technologies that developed in 1990 s have successfully probed the dynamics of single molecule enzyme catalysis in real time in vitro. Ever since then, single molecule enzymology has entered the golden age of rapid developing. Individual features of each enzyme hidden in the overall average have been discovered, and many new catalytic mechanisms have been proposed. Single molecule enzymology sheds light on the dynamic interactions between enzymes and substrates or products, deepening the understanding of biochemical reactions. This review described the recent research progresses of single molecule protease and ribozyme.

Objective To choose a reasonable non-drug treatment program for women with postpartum breast pain. Methods Based on an adequate assessment of the patients′ condition, the clinical questions were proposed and the references were searched in a series of databases, such as Cochrane Library, PubMed, Ovid, CINAHL, CNKI, Wanfang, Weipu, CBM. Results A preliminary search of 484 articles on cabbage therapy for postpartum breast pain was carried. Through rigorous preliminary screening and screening, 11 articles were finally included, including 2 systematic reviews, 2 randomized controlled trials and 7 quasi-experiment. Through the analysis of the inclusion literature, the data was extracted, and the evidence and summary evidence were strictly evaluated.According to the results of evidence, based on the patients′ condition and the wishes of the family, the cold and hot cabbage leaves were alternately applied to the breast of 10 postpartum women with breast engorgement, the breast distended pain were improved. Conclusions The method of evidence-based nursing can provide safe and effective treatment for postpartum women with breast engorgement.

Objective To discover more novel bat viruses and molecularly characterize bat-borne bocavirus diversity in Yunnan.Methods Twenty-six Aselliscus stoliczkanus were sampled in Jinghong , Yunnan, and subjected to viral metagenomic analysis.Specific PCR was used to detect any bocavirus in these samples based on the metagenomic result , while full genome was amplified and compared with other bocaviruses .Results and Conclusion Totally, 3 of the 26 (11.5%) bats were positive for bocavirus, the full genome of which contained 5203 nucleotides and could encode NS1, NP and VP1/VP2 proteins.Phylogenetic analysis showed that this virus shared up to 58.7% and 53.3% amino acid identities with canine bocavirus 1 and canine minute virus .According to ICTV criteria (85%amino acid sequence identity ) on a new species of bocavirus , this virus could be a novel species within genus Bocaparvovirus .This study provides important data to better understand viral diversity in bats and to uncover the relationship between bocavirus and its hosts .

0bjective To construct the program of diet for adults with crohn′s disease. Methods Based on evidence, two rounds of consultation with experts using Delphi method were conducted to construct the program of diet for adults with crohn′s disease. Results The diet program consisted of 5 first-level entries,12 second-1evel entries and 33 third-level entries.The response rate of both rounds' questionnaire were 100%;the authority coefficients were 0.808 and 0.811,respectively;the coordination coefficients ranged from 0.149 to 0.212. Conclusions The Evidence-based diet program for adult with Crohn′s disease was reliable, the experts who participated were representative, had consistent opinions on items of each level for the program of diet for adults with crohn′s disease. Experts also had high levels of enthusiasm and authority, and could provide some degree guidance for diet nursing.

Objective:To evaluate whether early enteral nutrition could benefit patients with different degrees of shock by dynamic changes of intestinal biomarkers intestinal fatty acid-binding protein (I-FABP) and citrulline.Methods:(1) From February 2021 to February 2023, 133 target patients in the Intensive Care Unit of Suzhou Hospital Affiliated to Nanjing Medical University were enrolled. The observation period was 7 days after admission, and intestinal biomarkers were monitored three times: 24 hours after admission (D1), the third day after admission (D3), and the seventh day after admission (D7). (2) The enrolled patients were divided into two groups according to the dose of norepinephrine received within 48 hours after admission: safe dose group [receiving norepinephrine < 0.3 μg/(kg·min)] and hazardous dose group [receiving norepinephrine ≥0.3 μg/(kg·min)]. The safe dose group was given early enteral nutrition according to the guidelines, and the dangerous dose group was randomly (random number) given early enteral nutrition (EEN) and delayed enteral nutrition (DEN).(4)The dynamic changes of intestinal fatty acid binding protein and citrulline in three groups were observed; The differences of intestinal biomarkers at different time points of dangerous dose of EEN/DEN were compared. The survival time of EEN/DEN group within 28 days was recorded, and Kaplan-Meier survival curve was drawn. Univariate and multivariate regression analyses of 28-day mortality were performed for the included population.Results:(1) The baseline data, APACHEⅡ score, arterial blood lactic acid, AGI grade, feeding interruption, total feeding time within 7 days, and 28-day survival number were statistically different between safe dose EEN group and hazardous dose EEN group ( P < 0.05). Compared the baseline data of the dangerous dose EEN group and the dangerous dose DEN group, only the number of feeding interruptions was statistically different ( P < 0.05). (2) The trend of change in the safe dose EEN group was the same as that in the dangerous dose DEN group: I-FABP decreased significantly from D3 to D7 ( P < 0.05); Citrulline decreased from D1 to D3, but increased from D3 to D7 ( P < 0.05). In dangerous dose EEN group, I-FABP had no significant change during the monitoring period ( P > 0.05). Citrulline decreased significantly from D1 to D3 ( P < 0.05). The EEN/DEN ratio at dangerous dose was significantly different only for D7-I-FABP ( P < 0.05). Compared with the survival curve of EEN/DEN at risk dose, DEN could improve the early survival rate of critically ill patients at risk dose (Breslow test P = 0.0447). (4) Age( OR=1.069,95% CI: 1.002-1.140, P=0.044) was independent risk factor for 28-day death . BMI ( OR= 0.772, 95% CI: 0.604-0.987, P=0.039), no feeding interruption ( OR=0.044,95% CI: 0.004-0.455, P=0.009), total feeding time within 7 days ( OR=0.959, 95% CI: 0.923-0.997, P=0.036) were the protective factors. Conclusions:EEN at the safe dose and DEN at the dangerous dose can effectively reverse the necrosis of enterocyte and promote the recovery of enterocyte function. EEN is not a clear risk factor for death at 28 days, but it not only increases the incidence of feeding interruption, but also do not conduct the recovery of intestinal cell function from the perspective of intestinal biomarkers.

Objective:To investigate the efficacy of atomization with budesonide, salbutamol, and acetylcysteine in the adjuvant treatment of bronchopneumonia in children.Methods:Seventy-two children with bronchopneumonia admitted to Huaiyuan Jingtu Hospital from July 2021 to June 2022 were retrospectively included in this study. These children were divided into BS and BSY groups according to different treatment methods. Based on conventional treatment, the BS group was given atomization treatment with budesonide and salbutamol, and the BSY group was given atomization treatment with budesonide, salbutamol, and acetylcysteine. After two courses of treatment, clinical efficacy, duration to improvements in symptoms and signs, adverse drug reactions, and changes in serum C-reactive protein and procalcitonin levels after treatment relative to those before treatment were compared between the two groups. The optimal medication plan was investigated.Results:The total response rate in the BSY group was 91.67% (33 cases/36 cases), which was significantly higher than 72.22% (26/36) in the BS group ( χ2 = 4.59, P = 0.032). The incidence of adverse drug reactions in the BSY group was 11.11% (4/36), which was significantly lower than 19.44% (7/36) in the BS group ( χ2 = 0.96, P = 0.326). After treatment, the levels of C-reactive protein and procalcitonin in the BSY group were (5.86 ± 5.66) mg/L and (2.59 ± 0.74) μg/L, respectively, which were lower than (15.64 ± 5.85) mg/L and (4.71 ± 0.93) μg/L in the BS group ( t = 7.20, 10.70, both P < 0.001). The durations to the disappearance of symptoms and signs including fever, cough, lung rales, and X-ray lung shadow in the BSY group were significantly shorter compared with the BS group ( t = 11.85, 4.19, 2.72, 2.39, all P < 0.05). Conclusion:Atomization with budesonide, salbutamol, and acetylcysteine in combination for the adjuvant treatment of bronchopneumonia in children can quickly relieve the clinical symptoms of children, improve the lung signs, reduce the degree of inflammation, and has a remarkable therapeutic effect on bronchopneumonia in children.

Objective To investigate the effect and mechanism of colchicine on mouse liver cancer via Hippo sig-naling pathway.Methods The 6-week-old male C57BL/6J mice were randomly divided into 3 groups:diethylni-trosamine(DEN)/carbon tetrachloride(CCl4)/ethanol(C2H5OH)induced mouse liver cancer model and col-chicine(0.1 mg/kg)intervention were established in control group,model group and colchicine group.From week 1st to week 2nd,the model group and the colchicine group were intraperitoneally injected with 1.0％DEN once a week.From week 3rd to week 7th,20％CCl4 dissolved in olive oil solution(5 ml/kg)was intragastric ad-ministration twice a week.From week 8th to week 18th,20％CC14 dissolved in olive oil solution(6 ml/kg)was intragastric twice a week.The colchicine group was given continuous intragastric administration for 20 weeks.The control group was given the corresponding solvent.Liver index,alanine aminotransferase(ALT)and aspartate ami-notransferase(AST)serum biochemical indexes were detected.Western blot and immunofluorescence were used to detect the expression levels of MST1,pYAP,YAP,pTAZ and TAZ proteins in liver tissues of mice in each group.Results The liver surface of mice in the control group was smooth and soft,while the liver of mice in the model group was rough and hard with granular nodules.The above lesions were significantly improved in the colchicine group.HE staining showed that the liver lobular structure of mice in the control group was normal,while the liver lobular structure of mice in the model group was disordered,with a small amount of fat droplets,extensive tissue necrosis,inflammatory cell infiltration,and fat vacuoles.The degree of liver lesions of mice after colchicine inter-vention was significantly reduced.The results of immunofluorescence and Western blot showed that compared with the control group,the protein expression levels of pYAP and pTAZ in liver tissue of model group mice were signifi-cantly decreased,while the protein expression levels of MST1,YAP and TAZ increased.After colchicine interven-tion,the protein expression levels of MST1,pYAP and pTAZ were significantly up-regulated,while the protein ex-pression levels of YAP and TAZ were down-regulated.Conclusion The therapeutic effect of colchicine on mouse liver cancer may be related to its activated Hippo signaling pathway.