Objective:To investigate the clinical effect of Paris polyphylla var. yunnanensis oral care solution in the prevention of ventilator-associated pneumonia (VAP) in patients with endotracheal intubation in ICU, and to provide reference for continuing to explore the best oral care solution of traditional Chinese medicine for preventing VAP. Methods:Randomized controlled experiment was used. Fifty patients with tracheal intubation admitted to the ICU of the Affiliated Hospital of Yunnan University from May 2022 to February 2023 were selected by convenient sampling method. The patients were divided into control group and observation group with 25 cases in each group by random number table method. The control group was treated with Xipayi mouth rinse for oral care, and the observation group was treated with self-made Paris polyphylla var. yunnanensis oral care solution for oral care. The incidence of VAP, clinical efficacy of oral care, leukocyte and procalcitonin after 7 days of intervention were observed, the mechanical ventilation time and ICU hospitalization time were recorded in the two groups. Results:All patients completed the study. There were 19 males and 6 females in the control group with the age of (60.72±19.32) years old. There were 15 males and 10 females in the observation group with the age of (60.24±16.80) years old. The incidence of VAP after 7 days of intervention in the observation group was 12% (3/25), which was lower than that in the control group 36% (9/25). The difference between the two groups was statistically significant ( χ2=3.95, P<0.05). The total clinical effective rate of oral care after 7 days of intervention in the observation group was 88% (22/25), which was higher than that in the control group 68% (17/25). The difference between the two groups was statistically significant ( χ2=9.78, P<0.01). There was no significant difference in the level of the leucocyte and procalcitonin before and after 7 days of intervention between the two groups (all P>0.05). The mechanical ventilation time and ICU hospitalization time in the observation group were (6.92 ±3.89), (10.36±6.17) days respectively, which were lower than those in the control group (9.72±4.76), (12.12±4.87) days. The difference of mechanical ventilation time between the two groups was statistically significant (t=2.28, P<0.05). The ICU hospitalization time in the observation group was lower than that in the control group, but there was no significant difference between the two groups ( P>0.05). Conclusions:Paris polyphylla var. yunnanensis oral care solution can improve the oral condition of patients, reduce the incidence of VAP, shorten the time of mechanical ventilation, which has certain clinical value.

Objective:To investigate the clinical effect of Paris polyphylla var. yunnanensis oral care solution in the prevention of ventilator-associated pneumonia (VAP) in patients with endotracheal intubation in ICU, and to provide reference for continuing to explore the best oral care solution of traditional Chinese medicine for preventing VAP. Methods:Randomized controlled experiment was used. Fifty patients with tracheal intubation admitted to the ICU of the Affiliated Hospital of Yunnan University from May 2022 to February 2023 were selected by convenient sampling method. The patients were divided into control group and observation group with 25 cases in each group by random number table method. The control group was treated with Xipayi mouth rinse for oral care, and the observation group was treated with self-made Paris polyphylla var. yunnanensis oral care solution for oral care. The incidence of VAP, clinical efficacy of oral care, leukocyte and procalcitonin after 7 days of intervention were observed, the mechanical ventilation time and ICU hospitalization time were recorded in the two groups. Results:All patients completed the study. There were 19 males and 6 females in the control group with the age of (60.72±19.32) years old. There were 15 males and 10 females in the observation group with the age of (60.24±16.80) years old. The incidence of VAP after 7 days of intervention in the observation group was 12% (3/25), which was lower than that in the control group 36% (9/25). The difference between the two groups was statistically significant ( χ2=3.95, P<0.05). The total clinical effective rate of oral care after 7 days of intervention in the observation group was 88% (22/25), which was higher than that in the control group 68% (17/25). The difference between the two groups was statistically significant ( χ2=9.78, P<0.01). There was no significant difference in the level of the leucocyte and procalcitonin before and after 7 days of intervention between the two groups (all P>0.05). The mechanical ventilation time and ICU hospitalization time in the observation group were (6.92 ±3.89), (10.36±6.17) days respectively, which were lower than those in the control group (9.72±4.76), (12.12±4.87) days. The difference of mechanical ventilation time between the two groups was statistically significant (t=2.28, P<0.05). The ICU hospitalization time in the observation group was lower than that in the control group, but there was no significant difference between the two groups ( P>0.05). Conclusions:Paris polyphylla var. yunnanensis oral care solution can improve the oral condition of patients, reduce the incidence of VAP, shorten the time of mechanical ventilation, which has certain clinical value.

OBJECTIVETo explore the effects of direct current electric fields on directional migration and arrangement of dermal fibroblasts in neonatal BALB/c mice and the related mechanisms.

METHODSTwelve neonatal BALB/c mice were divided into 4 batches. The skin on the back of 3 neonatal mice in each batch was obtained to culture fibroblasts. Fibroblasts of the second passage were inoculated in 27 square cover slips with the concentration of 5 × 10(4) cells per mL. (1) Experiment 1. Six square cover slips inoculated with fibroblasts of the second passage were divided into electric field group (EF) and sham electric field group (SEF), with 3 cover slips in each group. The cover slips were put in live cell imaging workstation. The cells in group EF was treated with electric power with EF intensity of 200 mV/mm, while simulating process without actual power was given to SEF group (the same below) for 6 h. Cell proliferation rate was subsequently counted. (2) Experiment 2. Six cover slips were divided and underwent the same processes as in experiment 1. Cell movement locus within EF hour (EFH) 6, direction change of cell migration at EFH 0 (immediately), 1, 2, 3, 4, 5, and 6 which was denoted as cos(α), cell migration velocity within EFH 6, direction change of long axis of cell within EFH 6, and direction change of cell arrangement at EFH 0, 1, 2, 3, 4, 5, and 6 which was denoted as polarity value cos[2(θ-90)] were observed under live cell imaging workstation. After EFH 6, the morphological changes in microtubules and microfilaments were observed with immunofluorescent staining. (3) Experiment 3. Six cover slips were divided into cytochalasin D group (treated with 1 μmol/L cytochalasin D for 10 min) and colchicine group (treated with 5 μmol/L colchicine for 10 min), with 3 cover slips in each group. The morphological changes in microfilaments and microtubules were observed with the same method as in experiment 2. (4) Experiment 4. Nine cover slips were divided into control group (no reagent was added), cytochalasin D group and colchicine group (added with the same reagents as in experiment 3), with 3 cover slips in each group. Cells in the 3 groups were exposed to an EF of 200 mV/mm for 6 h. Cell movement locus within EFH 6, cell migration velocity within EFH 6, cell polarity values at EFH 0, 3, and 6, and morphological changes of cells at EFH 0 and 6 were observed. Data were processed with independent samples t-test, one-way analysis of variance, and LSD test.

RESULTS(1) There was no statistically significant difference in cell proliferation rate in group EF and group SEF (t=-0.24, P﹥0.05). (2) Within EFH 6, cells in group EF migrated towards the anode of EF, while cells in group SEF moved randomly. At EFH 0, the values of cos(α) of cells in the 2 groups were both 0. The absolute value of cos(α) of cells in group EF (-0.57 ± 0.06) was significantly higher than that in group SEF (0.13 ± 0.09, t=6.68, P<0.01) at EFH 1, and it was still higher than that in group SEF from EFH 2 to 6 (with t values from 5.33 to 6.83, P values below 0.01). Within EFH 6, migration velocity of cells in group EF was (0.308 ± 0.019) μm/min, which was significantly higher than that in group SEF [(0.228 ± 0.021) μm/min, t=-2.76, P<0.01]. Within EFH 6, long axis of cells in group EF was perpendicular to the direction of EF, while arrangement of cells in group SEF was irregular. Cell polarity values in group EF were significantly higher than that in group SEF from EFH 2 to 6 (with t values from -7.52 to -0.90, P values below 0.01). At EFH 6, the morphology of microfilaments and microtubules of cells in EF group was similar to that in SEF group. (3) The fluorescent intensity of microfilaments of cells in cytochalasin D group became weakened, and the filamentary structure became fuzzy. The microtubules of cells in colchicine group became fuzzy with low fluorescent intensity. (4) Within EFH 6, cells in control group migrated towards the anode of EF, while cells in cytochalasin D group and colchicine group moved randomly. Within EFH 6, there was statistically significant difference in migration velocity of cells in the 3 groups (F=6.36, P<0.01). Migration velocity of cells in cytochalasin D group and colchicine group was significantly slower than that in control group (P<0.05 or P<0.01). At EFH 0, 3, and 6, cell polarity values in the 3 groups were close (with F values from 0.99 to 1.51, P values above 0.05). At EFH 0, cells in control group were spindle; cells in cytochalasin D group were polygonal or in irregular shapes; cells in colchicine group were serrated circle or oval. At EFH 6, no morphological change was observed in cells in control group; cells in cytochalasin D group were spindle with split ends on both ends; cells in colchicine group were serrated oval.

CONCLUSIONSThe physiologic strength of exogenous direct current EF can induce directional migration and alignment of dermal fibroblasts in neonatal BALB/c mice. Microfilaments and microtubules are necessary skeleton structure for cell directional migration induced by EF, while they are not necessary for cell directional arrangement induced by EF.

Animals ; Cell Movement ; Cells, Cultured ; Electricity ; Fibroblasts ; cytology ; Mice ; Mice, Inbred BALB C ; Microtubules ; Skin ; cytology

Immobilization of enzymes is important and widely applied in biocatalysis. Streptomyces platensis gene gox, encoding an extracellular L-glutamate oxidase (Gox), was fused to cellulose binding domain (CBDcex) from Cellulomonas fimi and the recombinant protein Gox-CBD was expressed in Escherichia coli. The fusion protein (Gox-CBD) was immobilized onto microcrystalline cellulose. The preparation conditions, binding capacity, properties and stability of the immobilized enzyme were studied. Under the condition of 4 ℃, for 1 hour, the fusion protein Gox-CBD was able to bind microcrystalline cellulose at a ratio of 9.0 mg of protein per gram of microcrystalline cellulose. Enzymatic properties of free and immobilized L-glutamic oxidase (Gox-CBD) were compared. The specific activity of the immobilized enzyme decreased, but its thermal stability increased a lot compared with that of the free Gox-CBD. After incubation at 60 ℃ for 30 min, 70% of the total activity remained whereas the free recombinant Gox completely lost its activity. The immobilized protein was tightly bound to microcrystalline cellulose at pH below 10 or more than 5 mmol/L NaCl. The fusion protein of Gox-CBD can be specifically immobilized on the microcrystalline cellulose on a single step. Therefore, our findings can provide a novel strategy for protein purification and enzyme immobilization.