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Corona Virus Disease 2019 (COVID-19) has seriously affected the treatment of patients and social stability. In the later stage of disease, some COVID-19 patients may develop into acute respiratory distress syndrome or even multiple organ failure. However, one of the most important mechanism underlying the deterioration of disease is cytokine storm. At present, some therapies such as interleukin-6 antibody blocker, stem cell therapy, and transfusion of convalescent plasma have been applied to counteract the cytokine storm and have made some progress. This article reviews the influences of cytokine storm syndrome on the COVID-19 and the corresponding immunotherapies to resist cytokine storm.
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Objective@#To explore the effects of dendritic epidermal T cells (DETC) on proliferation and apoptosis of epidermal cells in wound margin of mice and its effects on wound healing.@*Methods@#Twenty-eight healthy specific pathogen free (SPF) C57BL/6 wild-type (WT) male mice aged 8-12 weeks and 60 SPF T lymphocyte receptor δ-knockout (TCR δ-/-) male mice aged 8-12 weeks were selected to conduct the following experiments. (1) Eight WT mice were selected to isolate epidermal cells and primarily culture DETC according to the random number table. Morphological observation and purity identification of DETC by flow cytometer were detected immediately after culture and on culture day (CD) 15 and 30, respectively. (2) According to the random number table, 5 WT mice and 5 TCR δ-/- mice were selected and enrolled into WT control group and TCR δ-/- group. Round full-thickness skin defect with diameter of 6 mm was made on the back of each mouse. The wound healing condition was observed immediately after injury and on post injury day (PID) 2, 4, 6, 8, 10, and the percentage of residual wound area was calculated. (3) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, the tissue of wound margin was collected for hematoxylin eosin staining, and the length of new epithelium was measured. (4) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was collected to determine expression of proliferating cell nuclear antigen (PCNA) using Western blotting for evaluation of proliferation of epidermal cell. (5) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was selected and digested into single-cell suspension, and apoptosis of cells was detected by flow cytometer. (6) Forty TCR δ-/- mice were selected to carry out the same treatment as in experiments (2)-(5). According to the random number table, these mice were enrolled into TCR δ-/- control group and TCR δ-/-+ DETC group, with 5 mice in each group for each experiment. Round full-thickness skin defect was made on the back of each mouse. DETC in the number of 1×105 (dissolution in 100 μL phosphate with buffer purity above 90%) were injected through multiple points of wound margin of mice in TCR δ-/-+ DETC group immediately after injury, and equal volume of phosphate buffer was injected into mice of TCR δ-/- control group with the same method as above. Data were processed with one-way analysis of variance for repeated measurement, t test, and Bonferroni correction.@*Results@#(1) Along with the culture time elapse, the number of dendritic structures of DETC increased gradually. The percentage of T lymphocytes was 4.67% and 94.1% of these T lymphocytes were DETC. The purity of DETC on CD 15 was 18.50% and the purity of DETC on CD 30 was 98.70%. (2) Immediately after injury, the wound healing condition of mice in WT control group and TCR δ-/- group was similar. The wound healing speed of mice in TCR δ-/- group was slower than that in WT control group on PID 2-10. The percentages of residual wound area of mice in TCR δ-/- group on PID 2, 4, 6, 8, and 10 were increased significantly compared with those in WT control group (t=3.492, 4.425, 4.170, 4.780, 7.318, P<0.01). (3) The length of new epithelium of mice in TCR δ-/- group on PID 3 was (359 ± 15) μm, which was obviously shorter than that in WT control group [(462±26) μm, t=3.462, P<0.01]. (4) Immediately after injury, wound condition of mice in TCR δ-/-+ DETC group and TCR δ-/- control group was similar. Compared with TCR δ-/-+ DETC group, the wound healing speed of mice in TCR δ-/- control group were obviously slower on PID 2-10. The percentages of residual wound area of mice in TCR δ-/-+ DETC group on PID 2, 4, 6, 8, and 10 were decreased significantly compared with those in TCR δ-/- control group (t=2.308, 3.725, 2.698, 3.707, 6.093, P<0.05 or P<0.01). (5) On PID 3, the length of new epithelium of mice in TCR δ-/-+ DETC group was (465±31) μm, which was obviously longer than that in TCR δ-/- control group [(375±21) μm, t=2.390, P<0.05]. (6) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ-/- group was 1.25±0.04, which was obviously lower than that in WT control group (2.01±0.09, t=7.415, P<0.01). (7) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ-/-+ DETC group was 1.62±0.08, which was significantly higher than that in TCR δ-/- control group (1.05±0.14, t=3.561, P<0.05). (8) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ-/- group was (16.1±1.4)%, which was higher than that in WT control group [(8.1±0.6)%, t=5.363, P<0.01]. (9) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ-/-+ DETC group was (11.4±1.0)%, which was obviously lower than that in TCR δ-/- control group [(15.4±1.4)%, t=2.377, P<0.05].@*Conclusions@#DETC participates in the process of wound healing though promoting the proliferation of epidermal cells in wound margin and inhibit the apoptosis of these cells.
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Objective@#To explore effects of dendritic epidermal T cells (DETCs) and Vγ4 T lymphocytes on proliferation and differentiation of mice epidermal cells and the effects in wound healing of mice.@*Methods@#(1) Six C57BL/6 male mice aged 8 weeks were collected and divided into control group and wound group according to random number table (the same grouping method below), with 3 mice in each group. A 4 cm long straight excision with full-thickness skin defect was cut on back of each mouse in wound group, while mice in control group received no treatment. On post injury day (PID) 3, mice in 2 groups were sacrificed, and skin within 5 mm from the wound margin on back of mice in wound group and normal skin on corresponding part of mice in control group were collected to make single cell suspensions. The percentage of Vγ4 T lymphocyte expressing interleukin-17A (IL-17A) and percentage of DETCs expressing insulin-like growth factor Ⅰ (IGF-Ⅰ) were detected by flow cytometer. (2) Ten C57BL/6 male mice aged 8 weeks were collected and divided into control group and Vγ4 T lymphocyte depletion group with 5 mice in each group. Mice in Vγ4 T lymphocyte depletion group were injected with 200 g Vγ4 T lymphocyte monoclonal neutralizing antibody of Armenian hamster anti-mouse intraperitoneally, and mice in control group were injected with the same amount of Armenian hamster Ig intraperitoneally. One hole with full-thickness skin defect was made on each side of spine of back of each mice. The wound healing was observed on PID 1-8, and percentage of remaining wound area was calculated. (3) Six C57BL/6 male mice aged 8 weeks were grouped and treated in the same way as in experiment (2), with 3 mice in each group. On PID 3, expressions of IL-17A and IGF-Ⅰ in epidermis on margin of wound were detected with Western blotting. (4) Thirty C57BL/6 male mice aged 3 days were sacrificed, and epidermal cells were extracted. The keratin 14 positive cell rate was examined by flow cytometer (the same detecting method below). (5) Another batch of mouse epidermal cells were collected and divided into control group, IGF-Ⅰ group, and IL-17A group, with 3 wells in each group (the same well number below). Cells in IGF-Ⅰ group and IL-17A group were added with 1 mL recombinant mouse IGF-Ⅰ and IL-17A with final mass concentration of 100 ng/mL respectively, while cells in control group were added with the same amount of sterile phosphate buffered saline (PBS). On post culture day (PCD) 5, keratin 14 negative cell rate was examined. Another batch of mouse epidermal cells were collected, grouped, and treated in the same way as aforementioned experiment, and keratin 10 positive cell rate was examined on PCD 10. (6) Another batch of mouse epidermal cells were collected and added with 4 mmol/L 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) solution, and divided into control 0 d group, control 7 d group, IGF-Ⅰ group, and IL-17A group. Cells in IGF-Ⅰ group and IL-17A group were treated in the same way as the corresponding groups in experiment (5), and cells in control 0 d group and control 7 d group were treated in the same way as the control group in experiment (5). The CFSE fluorescence peaks were examined on PCD 0 of control 0 d group and PCD 7 of the other 3 groups. (7) Another batch of mouse epidermal cells were collected and divided into control group and IGF-Ⅰ group. Cells in IGF-Ⅰ group were added with 1 mL recombinant mouse IGF-Ⅰ with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, cells were underwent keratin 14 staining and CFSE staining as aforementioned, and keratin 14 negative cell rate of CFSE positive cells was examined. Another batch of mouse epidermal cells were collected and divided into control group and IL-17A group. Cells in IL-17A group were added with 1 mL recombinant mouse IL-17A with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, keratin 14 negative cell rate of CFSE positive cells was examined. Data were processed with one-way analysis of variance and t test.@*Results@#(1) On PID 3, percentage of DETC expressing IGF-Ⅰ in normal epidermis of control group was (9.9±0.8)%, significantly lower than (19.0±0.6)% of epidermis around margin of wound group (t=8.70, P<0.01); percentage of Vγ4 T lymphocyte expressing IL-17A in normal epidermis of control group was (0.123±0.024)%, significantly lower than (8.967±0.406)% of epidermis around margin of wound group (t=21.77, P<0.01). (2) On PID 1-4, there was obvious inflammatory reaction around wounds of mice in control group, and on PID 5-8, the wound area was still large. On PID 1-4, there was slight inflammatory reaction around wounds of mice in Vγ4 T lymphocyte depletion group, and on PID 5-8, the wound area was significantly reduced. On PID 3-7, percentages of residual wound area in Vγ4 T lymphocyte depletion group were significantly lower than those in control group (t=5.92, 5.74, 7.17, 5.38, 5.57, P<0.01), while percentages of residual wound area in two groups on PID 1, 2, 6 were similar (t=1.46, 3.17, 3.10, P>0.05). (3) On PID 3, compared with those in control group, expression of IL-17A and IGF-Ⅰ in epidermis around wound margin of mice in Vγ4 T lymphocyte depletion group was markedly decreased and increased respectively (t=8.47, 19.24, P<0.01). (4) The keratin 14 positive cell rate of mouse epidermal cells was 94.7%. (5) On PCD 5, the keratin 14 negative cell rate of mice in control group was markedly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=7.25, 5.64, P<0.01). On PCD 10, the keratin 10 positive cell rate of mice in control group was significantly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=3.99, 10.82, P<0.05 or P<0.01). (6) Compared with that of control 0 d group, CFSE fluorescence peaks of mouse epidermal cells in control 7 d group, IGF-Ⅰ group, and IL-17A group on PCD 7 shifted to the left. Compared with that of control 7 d group, CFSE fluorescence peaks of mouse epidermal cells in IGF-Ⅰ group and IL-17A group on PCD 7 shifted to the left. (7) On PCD 5, keratin 14 negative cell rate of CFSE positive cells of mice in control group was significantly higher than that in IGF-Ⅰ group (t=9.91, P<0.01), and keratin 14 negative cell rate of CFSE positive cells of mice in control group was markedly lower than that in IL-17A group (t=6.49, P<0.01).@*Conclusions@#In the process of wound healing, IGF-Ⅰ secreted by DETC can promote the proliferation of mouse keratin 14 positive epidermal cells and inhibit their terminal differentiation, while IL-17A secreted by Vγ4 T lymphocyte can promote the proliferation and terminal differentiation of mouse keratin 14 positive epidermal cells, thus both IGF-Ⅰ and IL-17A can affect wound healing.
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Objective To explore the effect of glucocorticoid combined with cyclophosphamide ( cytoxan, CTX) for the treatment of primary immunoglobulin A nephropathy ( IgAN ) with nephrotic syndrome and renal insufficiency .Methods 70 IgAN patients with nephrotic syndrome and renal insufficiency were selected as the research subjects,According to randomized single blind ,the patients were randomly divided into two groups ,35 cases in each group .The control group was treated with glucocorticoid ,the observation group was treated with glucocorticoid and cyclophosphamide combination therapy .The renal function indicators ,24 hour urine protein level ,serum albumin level,the occurrence of adverse reactions were compared between the two groups .Results After treatment, the endogenous creatinine clearance rate and glomerular filtration rate of the two groups were not decreased compared with before treatment (t =0.368,0.186,0.234,0.200,all P >0.05),the 24 hour urinary protein were decreased significantly (t=4.417,9.576,all P<0.05),serum albumin levels were significantly increased (t=5.922,9.433, all P<0.05).After treatment,the endogenous creatinine clearance rate [(16.23 ±7.57) mL/min,(16.09 ± 7.69)mL/min],glomerular filtration rate [(37.12 ±9.52) mL/min,(37.64 ±9.67) mL/min] between the two groups had no statistically significant differences (t=0.077,0.227,all P>0.05).After treatment,the 24 -hour urinary protein level and serum albumin level of the observation group [(1.18 ±0.30)g,(36.24 ±2.39)g/L]were better than those of the control group[(1.50 ±0.27)g,(33.92 ±2.07)g/L,t=4.691,4.341,all P<0.05].During the treatment, the incidence rate of adverse reactions in the observation group ( 25.71％) had no statistically significant difference compred with that in the control group (22.86％) (χ2 =0.078,P>0.05).Conclusion For IgAN patients with nephrotic syndrome and renal insufficiency , glucocorticoid combined with cyclophosphamide can reduce the urine protein ,increase serum albumin concentration to a certain extent ,but it has no obvious effect on renal function,and the adverse reaction is more serious than the single use of glucocorticoids .
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Objective To study conventional laparoscopic instruments line single hole peritoneoscope gallbladder excision and three hole laparoscopic cholecystectomy surgery efficacy and safety.Methods 140 cases patients with gallbladder stones in our hospital from January 2014 to June 2015 were selected as the research subjects.All patients need to be treated with cholecystectomy.All the patients according to the random number table were randomly divided into two groups,respectively for single hole group and three group.Single hole group using a single hole peritoneoscope gallbladder excision,triplex group use three hole laparoscopic gallbladder resection.The surgical effect,hospitalization costs and complications were compared between the two groups after treatment.Results Hole group operation time (87.89 ± 12.81) min,longer than the three-hole group (53.89 ± 8.91) min,but the hospital stay was (1.28 ± 0.21) d and total hospital costs (11 241.21 ± 23.91) yuan,were lower than three-hole group,P ＜ 0.05,the difference was statistical significance in the amount of bleeding in the two groups were not statistical significance,P ＞ 0.05;two groups of patients had complicatiom,but have been treated better,and two concurrent disease (1.43％ vs 2.86％) incidence was no significant difference,P ＞ 0.05.Conclusion Single hole laparoscopic cholecystectomy reduce the hospitalization time and hospitalization expenses,surgical trauma is smaller,postoperative recovery is faster,and the safety is high,it is worthy of clinical application.
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To explore the role of dentritic epidermal T lymphocytes ( DETCs) in immune rejection of skin allograft in mice and its related mechanism. Methods (1) Full-thickness skin was harvested from back of one male wild type (WT) C57BL/6 mouse. Epithelial cells were isolated for detection of the expression of DETCs and their phenotype with flow cytometer. Another male WT C57BL/6 mouse was used to harvest full-thickness skin from the back. Epidermis was isolated for observation of the morphological characteristics of DETCs with immunofluorescence technology. (2) Four male green fluorescence protein (GFP)-marked C57BL/6 mice, 7 female WT C57BL/6 mice (group WT), and 7 female ybT lymphocytes 8 gene knock-out (GK) C57BL/6 mice (group GK) were used. Full-thickness skin in the size of 1.4 cm x 1.4 cm on the back of mice in groups WT and GK were excised, and the wounds were transplanted with full-thickness skin in the size of 1.2 cm x 1.2 cm obtained from male GFP-marked C57BL/6 mice. The survival time of skin grafts was affirmed with small animal in vivo imager and naked eyes and recorded. (3) Two male WT C57BL/6 mice were used to isolate epithelial cells. Cells were inoculated into 48-well plate and divided into activation group (A) and control group (C) according to the random number table, with 4 wells in each group. Cells in group A were treated with 10 pL concanavalin A in the concentration of 2 microg/mL for 24 hours, while those in group C with PBS in the same volume as that in group A. The expression of interferon y in DETCs was detected with flow cytometer. (4) Four male GFP-marked C57BL/6 mice were used as donors. Fourteen female WT C57BL/6 mice were used as receptors and divided into interferon gamma neutralizing group (IN) and control group (C) according to the random number table, with 7 mice in each group. The skin transplantation model of C57BL/6 male to C57BL/6 female was established as in part (2). Before surgery and 72 hours after, mice in group IN were intraperitoneally injected with 200 pL interferon y neutralizing antibody in the concentration of 1 mg/mL, and those in group C with normal saline in the same volume as that in group IN. The survival time of skin grafts was observed and recorded using the methods in part (2), and the result of group IN was compared with that of group GK in part (2). The survival curve of skin grafts was processed with Log-rank ( Mantel-Cox) test. Results (1) The positive expression rate of DETCs in epithelial cells of skin in mouse was 7.27%, and they were all CD3 cells. DETCs were found to be scattered in the epidermis of skin in mouse with dendritic morphology. (2) The survival time of skin grafts of mice in group GK was 22-35 d, obviously longer than that in group WT (12-16 d, y2 = 14. 10 , P < 0.001). (3) Expression of interferon gamma was detected in 22. 70% DETCs in group A, which was obviously higher than that in group C (0.51%). (4) The survival time of skin grafts of mice in group IN was 19-24 d, which was obviously longer than that in group C (12-16 d, chi 2 = 13.60, P < 0.001) but close to that in group GK as in part (2) (chi2 = 0.06, P = 0.810). Conclusions DETCs are involved in promotion of immune rejection of skin allograft probably by secretinf interferon gamma.
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Animals , Female , Male , Mice , Allografts , Epidermis , Graft Survival , Allergy and Immunology , Physiology , Interferon-gamma , Allergy and Immunology , Metabolism , Lymphocytes , Mice, Inbred C57BL , Skin , Skin Transplantation , T-Lymphocytes , Allergy and ImmunologyABSTRACT
Objective To analyze the clinical results of modified N-butyl 2-cyanoacrylate skin adhesive in the application of facial trauma.Methods A total of 687 patients had been analyzed retrospectively from August 2009 to July 2014,and all the wounds of these patients had not obtained the anatomical repositioning after pure manual reduction with tissue glue or pure stitches.Improved tissue adhesive method was as follows:point coating,decomposition and glue with stitches.Results Anatomical repositioning was obtained in 685 wounds of 687 cases with improved tissue adhesive method.Infection occurred in the wounds of 2 cases (0.29％),but no other complications occurred in all patients.During postoperative 3 months of follow-up,clinical results were satisfactory for all patients.Conclusions The improved method of tissue glue has fewer postoperative complications,beautiful and more satisfatory appearance,economic,safer and reliable in the application of facial trauma.
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Background Corneal walleye formation is a result of excessive corneal wound healing.Controlling the corneal trauma or excessive healing reaction after operation of penetrating corneal trauma is a focus in the study of corneal wound healing.Objective This study was to investigate the changes of corneal strength and the expression of keratocan,a marker of corneal stroma,after corneal penetrating injury operation,and to determine the optimal removal time of corneal suture.Methods Full-thickness incisions of 5 mm along the vertical diameter were done at the central cornea on 80 eyes of 80 six-month-old New Zealand rabbits in this study.Then the incisions were interruptedly sutured with 10-0 nylon thread to establish the corneal wound healing models.Each 4 rabbits were sacrificed in 1 week,2,3,4,5,6,7,8 weeks after operation,respectively,and the central corneal stripes were prepared with the size of 7 mm×5 mm.The mean maximal strength of the corneal bands was measured by uniaxial tensile test with electroforce3220-AT biomachanics machine.Then,each 6 whole corneas were obtained at the above time points,and the dynamic changes of keratocan mRNA expression in the specimens were detected by reverse trancription PCR (RT-PCR).The care and use of the animals followed the rules of ARVO.Results The mean maximal strength of corneal stripes was 0,(1.007 ± 0.041),(1.991 ± 0.034),(2.512 ± 0.030),(3.630 ± 0.049) and (4.935 ± 0.004)M Pa in 1 week,2,3,4,5,6 weeks after modeling,and the corneal strength values from 3 weeks through 6 weeks were significantly enhanced in comparison with the value at the adjacent before timepoint (q =6.35,7.54,8.21,5.86,all at P＜0.01).The relative expression levels (absorbance) of keratocan mRNA in the corneas were 0.869±0.015,0.779 ± 0.065,0.621 ± 0.027,0.460 ± 0.018,0.393 ± 0.057 and 0.255 ± 0.045 in 1 week,2,3,4,5,6 weeks after operation,and each value was lower than that of the adjacent before timepoint (q =5.24,5.61,7.49,4.75,5.47,all at P＜0.01).The intensity of corneal stripes and the expression levels of keratocan mRNA in the corneas were stable in 7 weeks and 8 weeks after operation.Conclusions The dynamic change of corneal strength during the repair of penetrating corneal injury is associated with the down-regulation of keratocan in cornea.Rabbit cornea reaches a maximal strength capacity in 6 weeks after penetrating injury,therefore,it is the optimal time to remove suture.
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<p><b>OBJECTIVE</b>To construct a 3D finite-element model of the craniofacial complex with the original DICOM data of CT and to investigate the preliminary biomechanical characteristics with different directions and magnitudes of retractive forces to the maxilla of rhesus monkeys.</p><p><b>METHODS</b>A male rhesus monkey with mixed dentition was used. Spiral CT was performed to establish a 3D finite-element model of the craniofacial complex. The ANSYS 12.1 software was used to analyze craniofacial complex displacement.</p><p><b>RESULTS</b>Each landmark showed larger displacement with increasing force value. The displacement values and force size exhibited a linear relationship. In the x-axis direction, all displacements were small. In the y-axis direction, all displacements showed significantly higher changes with increasing force value displacement. In the z-axis direction, the A-point and ANS point moved downward, but PNS moved upward.</p><p><b>CONCLUSION</b>Loading retractive force resultes in an apparent backward and clockwise rotation on the maxilla with no obvious effects on the width of the upper jaw.</p>
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Animals , Male , Biomechanical Phenomena , Dental Occlusion , Dental Stress Analysis , Macaca mulatta , MaxillaABSTRACT
Objective To evaluate the effect of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B (UVB)-induced premature senescence and photocarcinogenesis in human skin fibroblasts (HSFs).Methods A previously established HSF cell clone with repressed expression of p53,which was named as HSF-p53,was cultured and irradiated with a subcytotoxic dose (10 mJ/cm2) of UVB once a day for five consecutive days.The HSFs with normal expression of p53 served as the control.Subsequently,β-galactosidase (SA-β-gal)-staining was performed to estimate the degree of senescence,quantitative real-time PCR array was performed to determine the mRNA expressions of photocarcinogenesis-and senescence-associated genes,including p53,p21,p19,p16,pRb,fibronectin,osteonectin,smooth muscle 22 (SM22),bax,bcl-2,hypoxia-inducible factor-1 α (HIF-1α),vascular endothelial growth factor(VEGF),and human double minute-2 (hdm2).Statistical analysis was carried out by Student's t test using the software SPSS 10.0.Results The percentage of SA-β-gal-positive cells in irradiated HSF-p53 was 19.70％ ± 0.85％,significantly higher than that in unirradiated HSF-p53 (12.77％ ± 0.81％,t =6.45,P ＜ 0.05),but lower than that in irradiated control HSFs (50.48％ ± 5.30％,t =7.86,P ＜ 0.05),and similar to that in unirradiated control HSFs (18.50％ ± 0.45％,t =2.57,P ＞ 0.05).Compared with the control HSFs,the HSF-p53 showed decreased expressions of p21,p19,fibronectin,osteonectin,SM22 and bax genes (all P ＜ 0.05),but increased expressions of bcl-2,HIF-1α,VEGF and hdm2 genes (all P ＜ 0.05),and a similar expression of p16 gene (P ＞ 0.05); the repeated UVB radiation significantly promoted the expressions of p16 and pRb genes (both P ＜ 0.05),but had no obvious effect on the expressions of the other genes in HSF-p53 compared with unirradiated HSF-p53 (all P ＞ 0.05).Conclusions The inhibition of p53 expression may decelerate the UVB-induced premature senescence in HSFs,which may be involved in the p53-dependent tumor suppression.
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Objective To investigate the clinical value of 64-slice spiral computed tomography(64-MSCT)triple-phase enhanced scan in diagnosis of lymphatic metastasis of gastric cancer.Methods Thirty patients with gastric cancer underwent plain and triple-phase enhanced scan by using 64-MSCT to analyze the relevant parameters of lymphatic metastasis.Results The four parameters de-termined metastatic perigastric lymph node as follows:①the short diameter ≥6 mm,②the ratio of short-to-long diameter ≥0.6,③the CT value in the portal venous phase≥ 65 HU,④the difference of CT values between portal venous phase and plain scan≥35 HU.The sensitivity and specificity of combining two parameters (①+②)in diagnosing metastatic lymph node were 90.5% and 29.0%,respectively.The sensitivity and specificity of combining three parameters (①+②+③)were 98.2% and 1 9.4%,respec-tively.The sensitivity and specificity of combining four parameters (①+②+③+④)were 99.7% and 13.2%,respectively.In ad-dition,metastatic lymph nodes were considered if they were ring-enhancement,or adhesions of several lymph nodes.Conclusion The use of 64-MSCT triple-phase enhanced scan and synthesis of various parameters of lymph nodes could lead to reliable diagnosis of lymphatic metastasis in gastric cancer with rapid,non-invasive,high sensitive and specific features.
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Objective To investigate the effect of employing achievement management in nursing management of urology department.Method The results of assessments were linked to the achievements by regulating achievement-oriented distribution and assessment methods.Results The degree in patients' satisfaction with nursing service and the score on nursing quality assessment were both significantly improved compared to pre-performance of achievement-oriented distribution and assessment(P<0.01). Conclusion The achievement-oriented management may be effective in mobilizing nurses' working enthusiasm and increase quality of care,which guarantees fundamental nursing care.
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To construct, express, purify and identify the Myc-R9-EGFP fusion protein and validate its transduction activity in the cultured porcine embryo fibroblasts. cDNA of pig c-Myc gene was amplified by RT-PCR with specific primers of 9 arginine (R9) from the primordial genital ridges and inserted into prokaryotic expression vector pET-28a-EGFP. After DNA sequencing confirmation, the recombinant plasmid was then transformed into BL21 (Escherichia coli) strain. After IPTG induction, the target fusion protein was efficiently induced to express, successfully purified by Novagen His-Bind kit, identified by SDS-PAGE and Western blotting. Finally, its high transduction activity in the porcine embryo fibroblasts was validated. The purified Myc-R9-EGFP fusion protein and the validation of its transduction activity in fibroblasts have provided an experimental foundation for further studies on the biological characterization of Myc protein, and soundly facilitated the further study of establishing pig induced pluripotent stem cells by recombinant protein.
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Animals , Arginine , Genetics , Metabolism , Cell Line , Cell Membrane Permeability , Escherichia coli , Genetics , Metabolism , Fibroblasts , Cell Biology , Metabolism , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Proto-Oncogene Proteins c-myc , Genetics , Metabolism , Recombinant Fusion Proteins , Genetics , Metabolism , Pharmacokinetics , SwineABSTRACT
Objective To establish the immunization model with synthetic M3-muscarinic receptor peptides in rats and investigate the role of autoantibodies against M3-muscarinic receptor in the pathogenesis of chronic obstructive pulmonary disease. Methods The control and immunization models were established. The rate of positive of autoantibodies against M3-muscarinic receptor and the pulmonary function were detected. The blood gas analysis was also detected on the next day after finishing immunization. The tissues of lung were observed by light microscope. Results In immunization group, the rate of positive of autoantibodies against M3-muscarinic receptor were 100%, the geometric means of autoantibody were 1:152. There was a statistical difference between the immunization group and control group (x2 =6. 68, P ＜0. 01). The inspiratory resistance (Ri) was [ ( 1. 77 ±0. 22) cm H2O/ml. sec] and [ ( 1.39±0. 21 )cmH2O/ml. sec] in immunization and control group, respectively. It was increased while the lung compliance (C1) and the percentage of forced expiratory volume in first 0. 3 second to forced vital capacity ( FEV0. 3/FVC% ) were decreased in immunization group. There was a statistical difference compared with control group ( t = 3. 11,2. 82,3.23, allP ＜ 0. 01 ). In immunization group, the PaCO2 was higher and the pH as well as the PaO2 were lower than that in control group. The values of blood gas analysis showed a statistical difference between the immunization group and control group ( t =3.86,3. 47 ,allP ＜0. 01 ). Lung tissueswere severely destroyed in immunization group. There were many inflammation cells and hyperplasia glands,alveolar septum was very thick, and part of pulmonary alveoli even expanded to form bullae lung. Positive correction were found between autoantibodies against M3-muscarinic receptor and pulmonary function as well as blood gas analysis in immunization group. Conclusion The autoantibodies against M3-muscarinic receptor maybe play an important role in the pathogenesis of chronic obstructive pulmonary disease.
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Objective Idiopathic pulmonary fibrosis (IPF) and nonspecific interstitial pneumonia (NSIP) are the two largest subsets of idiopathic interstitial pneumonia (IIP). They have a sharply differences in therapy and survival, however, the identification of them is difficult In general, NSIP has a much better responsiveness to therapy and good survival On the other hand, in IPF, was demonstrated to be associated with poorer survival and responsiveness to therapy. To diagnosis of IPF and NSIP need histopathologic classification which however, requires surgical lung biopsy. Therefore, we try to set up an equation using those data from non-invasive methods to identify IPF and NSIP. Methods A retrospective review of 32 patients with IPF (n = 14) and NSIP (n = 18) was carried out. General clinical data, pulmonary function, chest radiographic, and bronchoalveolar lavage fluid were recorded. The statistical methods of logistic regression and multiple linear regression are used to establish the equation. The method of curve fitting is used to find the critical value of the equation to the differential diagnosis between the IPF and NSIE Results (1) Compare to NSIP, patients of IPF are older, more males than females, having a bigger proportion of smokers. (2) Compared with NSIP patients, IPF patients have higher CT score of "grid shadow", "cellular shadow" and lower CT score of "ground-glass shadow". (3) Bronchoalveolar lavage (BAL) lymphocytosis was more frequently observed in NSIP than IPF. (4) We get an equation: y = 0. 9 + 0. 123x1 -0. 045x2 +0. 009x3 +0. 033x4 and it can improve the differential diagnosis between the IPF and NSIP in this group of patients. Conclusion Clinical,high resolution computerized tomography and BAL are useful non-invasive tools in diagnosis IPF and NSIP. The equation: y = 0.9 + 0. 123x1 - 0.045x2 + 0. 009x3 +0. 033x4 can help us to distinguish the IPF and NSIP.
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From 2006 to 2008, the susceptibility of different species of animal hosts to Campyebacter jejubni infection was observed in various areas of Jiangsu province, in which the API Campy System was used to perform the biochemical identification and the multiple PCR assay was employed to analyse the C.jejuni isolates from 3010 specimens of fouls. Cattle, pigs and monkeys, and in addition, the susceptibility of isolates to various antibiotics was also determined. In these specimens investigated 402 samples were found to be positive in the detection of C.jejuni with a positive detection rate of 13.36%. The positive detection rates in chicken, water fouls, milk cows, pigs, monkey, red crowned crane and wapiti were 15.83% (258/1630), 10.4% (52/100), 8.24% (42/510), 15.63% (25/160), 15% (15/100), 12.5% (15/80) and 0% (0/30) respectively. Meanwhile, the antibiotics to which the isolates from different hosts showed high rate of sensitivity to 27 antibiotics of 10 varieties included: chloromycetin (100%), Almocylin (99.7%), amicarcin (92.59%), cefprozil (91.67%), alchimycin (90.74%); while the antibiotics to which these isolates showed high rate of resistance were compound neoromin (99.7%), cefoperazone (99.07%), trimethoprim (97.22%), cepronatin (91.67%), cepromondo (99,07%) respectively. It is evident that the susceptibility of different hosts to C.jejuni infection and the status of drug-resistance of the isolates appear to be quite different and more complicated in Jiangsu province.
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To improve the composit application knowledge and clinic performance ability of the advanced study doctor.We take the clinc doctors as study object,using various teaching ways to discuss the methods and quality of orthodontic doctors.Through making rigorou teaching planing,seting reasonable courses,and strict clinic operational training,the quality of teaching is visible improved.This teaching system is worth to make use and extend.
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Objective To screen the proteins interacting with FOXP3 in yeast two-hybrid system. Methods The "bait plasmid" pGBKT7 (named as pGBKT7-FOXP3) was constructed successfully. Using FOXP3 as bait, a human liver cDNA library was screened and the proteins interacting with FOXP3 were searched. The false positive clones were discarded by one to one yeast two-hybrid system, and the positive clones were sequenced and analyzed by bioinformatic methods. Results The bait plasmid pGBKT7-FOXP3 was constructed successfully and there was no self-activation or toxicity in AH109. Three proteins had been found in our system to be able to interact with FOXP3. They were tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein. Conclusion FOXP3 interacts with tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein, all of which may interfere in cell metabolism and function of T cell.
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Objective To express human HT036 protein in Escherichia coli(E.coli.)and identify it.Methods The cDNA sequence obtained by PCR was cloned into the prokaryotic expression plasmid pET30a(+).The target protein was expressed in E.coli..induced by IPTG and analyzed by Western blotting.Results The interest gene was identified by restriction endonucleases digestion and DNA sequencing.The protein was highly expressed in E.coli..Conclusion We successfully expressed the HT036 protein.
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Objective To explore the interaction between HT036(hypothetical protein HT036)and P311 by co-immunoprecipitation.Methods HA-tagged fusion protein(HA-HT036)expression vector was constructed,identified and transfected into human embryo kidney 293(HEK293)cells alone or with Myc-tagged fusion protein(Myc-P311)expression vector pCMV-Myc-p311.The interaction between P311 and HT036 was detected by co-immunoprecipitation.Results Double restriction enzyme digestion showed that pCMV-HA-HT036 was constructed correctly.When Myc-P311 was immunoprecipitated by anti-Myc antibody,HA-HT036 was identified by Western blotting with anti-HA antibody from immunoprecipitated complex.Conclusion The recombinant vector pCMV-HA-HT036 was constructed successfully.The interaction between HT036 and P311 could be identified by co-immunoprecipitation after co-expression of pCMV-HA-HT036 and pCMV-Myc-p311.The result provides an important basis for further study of the intracellular signal transduction of P311.