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ObjectiveTo establish an allele-specific polymerase chain reaction （PCR） method for identifying Scolopendra dispensing granules， so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3（COX-3） gene sequence of Scolopendra， and the single nucleotide polymorphism （SNP） loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature， cycles， Taq enzymes， DNA template amount， PCR instruments， and primer concentrations， and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix， 0.4 μL forward primer （10 μmol·L-1）， 0.4 μL reverse primer （10 μmol·L-1）， 2.5 μL DNA template， and 9.2 μL sterile double distilled water. PCR parameters： Pre-denaturation at 94 ℃ for 3 min， 30 cycles （94 ℃ for 20 s， 62 ℃ for 20 s， 72 ℃ for 45 s）， and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above， the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials， decoction pieces， and standard decoction freeze-dried powder of Scolopendra， as well as the intermediates and final products of Scolopendra dispensing granules， which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.
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In the quality control of Chinese medicine， the detection of active components and toxic and harmful components are two important links. Although conventional methods such as high performance liquid chromatography and liquid chromatography-mass spectrometry can accurately quantify the above substances， they have shortcomings such as complicated operation， high costs， inability of detection at any time， difficult detection of insoluble and macromolecular substances. Enzyme-linked immunosorbent assay （ELISA） can adsorb antigens or antibodies on the surface of solid carriers and realize qualitative or quantitative analysis of targets by using the specific reactions of antigens and antibodies. This method is praised for the simple operation， high sensitivity， strong specificity， simple requirements for experimental equipment， a wide application range， and low costs. In recent years， ELISA has been widely used in the quality control of Chinese medicine， especially in the content determination of mycotoxins represented by aflatoxin and the qualitative and quantitative analysis of active components. ELISA plays an increasingly important role with its unique advantages， providing new methods and ideas for the rapid quality examination of large quantities of Chinese medicines. This paper reviews the research progress in ELISA for the quality control of Chinese medicine in recent years and prospects its technical development and application prospects， aiming to provide reference and research ideas for further using this method to ensure the quality， safety， and controllability of Chinese medicine.
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@#This study was designed to investigate the therapeutic effect of anti-inflammatory tripeptide KdPT on ophthalmoxerosis. Male BALB/c mice, 8-week old, were treated with 0.2% benzalkonium chloride solution to establish the ophthalmoxerosis model. Four weeks after modeling, the mice were randomly divided into control group, positive group and the low, medium, high dose groups of KdPT. Each group was given normal saline, artificial tears and 1, 10, 100 μg/mL KdPT, respectively. After 3, 5, 7, 10 and 14 days of treatment, the morphology of the eye surface was observed, and the fluorescein sodium staining score was performed. The amount of tear secretion was measured by phenol red cotton thread and the right corneas were taken out for histopathological analysis after 14 days of treatment. Data showed that there was no significant abnormality in general state and the weight of mice in each group at each time point of treatment. After 14 days of treatment, KdPT can promote the secretion of tear, repair the damaged corneal epithelium, and showed a significant therapeutic effect on ophthalmoxerosis in mice. Based on the data, it is possible for KdPT to be developed as a novel drug for ophthalmoxerosis.
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Objective@#Through the study of angiotensinⅡ- type 2 receptor agonist (AT2R) after pretreatment of mechanical ventilation lung injury (VILI) in rats model, to clarify the role of angiotensin Ⅱ - type 2 receptor agonist (C21) in alleviating VILI inflammation and the damage of immune function and its possible mechanism.@*Methods@#In this experiment, the acute lung injury model was established by mechanical spring-volume ventilation in SD rats, and C21 pretreatment was performed to observe the pathological condition of lung tissue in rats with different ventilation duration, and to detect the inflammatory changes of BALF lavage fluid. Flow cytometry was used to detect the CD68+/iNOS+ labeled M1 type AMφ and the CD68+/Arg-1+ labeled M2 type AMφ in alveolar lavage fluid.@*Results@#The mechanical VILI rat model was successfully established. The pathological injury score of the mechanical ventilation 4 h model, the wet/dry weight of lung tissue, the number of cells and protein in BALF lavage fluid were increased significantly, the levels of TNF-α and IL - 1 were increased significantly, the levels of IL-4 and IL-10 were decreased significantly, and the level of inflammatory reaction decreased with the increase of ventilation time. The M1/M2 ratio in the 4 h ventilation model group was the highest, which was significantly different from the control group (P<0.05). Compared to the model group, the C21 pretreatment significantly reduced the levels of proinflammatory factors TNF-αand IL-1, and increased the levels of anti-inflammatory factors IL-4 and IL-10 in BALF (P<0.01). After the intervention of AT2R agonist at 4 h, 6 h and 8 h, the M1/M2 ratio of the model was lower than that of the model without AT2R agonist at 4 h, 6 h and 8 h.@*Conclusion@#Mechanical ventilation for 4 h in SD rats can establish a mechanical ventilation lung injury model. AT2R agonist C21 can promote the polarization of macrophages to m2-type, and C21 pretreatment may alleviate VILI inflammation and immune damage by altering the polarization of macrophages.
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Objective Through the study of angiotensin Ⅱ-type 2 receptor agonist(AT2R)after pretreatment of mechanical ventilation lung injury(VILI)in rats model,to clarify the role of angiotensin Ⅱ-type 2 receptor agonist(C21)in alleviating VILI inflammation and the damage of immune function and its possible mechanism.Methods In this experiment,the acute lung injury model was established by mechanical spring-volume ventilation in SD rats,and C21 pretreatment was performed to observe the pathological condition of lung tissue in rats with different ventilation duration,and to detect the inflammatory changes of BALF lavage fluid.Flow cytometry was used to detect the CD68+/INOS+labeled M 1 type Amφ and the CD68+/Arg-1+labeled M2 type Amφ in alveolar lavage fluid.Results The mechanical VILI rat model was successfully established.The pathological injury score of the mechanical ventilation 4 h model,the wet/dry weight of lung tissue,the number of cells and protein in BALF lavage fluid were increased significantly,the levels of TNF-α and IL-1 were increased significantly,the levels of IL-4 and IL-10 were decreased significantly,and the level of inflammatory reaction decreased with the increase of ventilation time.The M1/M2 ratio in the 4 h ventilation model group was the highest,which was significantly different from the control group(P＜0.05).Compared to the model group,the C21 pretreatment significantly reduced the levels of proinflammatory factors TNF-α and IL-1,and increased the levels of anti-inflammatory factors IL-4 and IL-10 in BALF(P＜O.O1).After the intervention of AT2R agonist at 4 h,6 h and 8 h,the M1/M2 ratio of the model was lower than that of the model without AT2R agonist at 4 h,6 h and 8 h.Conclusion Mechanical ventilation for 4 h in SD rats can establish a mechanical ventilation lung injury model.AT2R agonist C21 can promote the polarization of macrophages to m2-type,and C21 pretreatment may alleviate VILI inflammation and immune damage by altering the polarization of macrophages.
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Aim To explore whether the polypeptide vaccines CKL9 and YL20 can induce immune response and anti-tumor effect on HER-2 (+) tumors in vitro and in vivo, and to provide suggestions for clinical use.Methods The proliferation of specific lymphocytes and cytotoxic T lymphocyte activity(CTL) stimulated by CKL9 and YL20 were studied with CCK-8 assay and LDH assay, and the antitumor activity of CKL9 and YL20 was evaluated in vivo.Results The lymphocyte proliferation was promoted by incubation with CKL9 and YL20, and the relative increase of cells was 11.1% and 16.7% respectively at the concentration of 50 mg·L-1 of CKL9 and YL20.The LDH assay confirmed the CTL effect induced by CKL9 and YL20 on HER2-positive tumor cells, not on HER2-negtive tumor cells.With an effector-target ratio of 80 ∶1, the inhibition of tumor cell by cytotoxic T lymphocyte stimulated by CKL9 and YL20 could reach 89.8% and 84.3%, respectively.The HER2(+) tumor cell N87 transplanted in Babes mice was inhibited by pre-immune polypeptide CKL9 and YL20.Conclusion The HER2-specific polypeptide vaccines CKL9 and YL20 could induce persistent specific CD4 and CD8 T cell immune and inhibit the growth of HER2 positive tumor cells.
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Objective To investigate the protective effects of Platycodon grandiflorum total saponins (PGTS) on acute lung injury (ALI) in rats and the related mechanisms.Methods Total of 60 SD rats were randomly (random number) divided into control group,model group,low-,middle-and high-dose PGTS group,and dexamethasone group,10 rats in each group.The latter 4 groups and dexamethasone group were injected with 50,100,200 mg/kg PGTS and 5 mg/kg dexamethasone,respectively.After 1 h,the latter 5 groups were intraperitoneally injected with mg/kg LPS to establish the ALI model.The clinical symptoms of the rats were observed.After 12 h,the arterial PaO2 and PaCO2,serum TNF-α and IL-10 level,lung wet/dry weight ratio (W/D),lung tissue SOD,GSH-Px and MDA level and NF-κB protein expression were determined.Results Rats in model group manifested noticeable symptoms of acute lung injury (ALI) and lung tissue lesions.In treatment group with appropriate PGTS dose,ALI symptoms and lung lesions were significantly alleviated,arterial PaO2 was markedly increased (P ＜ 0.05),PaCO2 was decreased obviously (P ＜ 0.05),serum TNF-α level was prominently decreased (P ＜ 0.05),IL-10 level was strikingly decreased (P ＜ 0.05),lung W/D ratio was significantly decreased (P ＜0.05),lung tissue SOD and GSH-Px level were distincdy increased (P ＜0.05),MDA was clearly decreased (P ＜ 0.05),and NF-κB protein expression was plainly decreased (P ＜ 0.05),compared with model group.Conclusions PGTS has undoubted protective effects on acute lung injury induced by LPS in rats.The mechanism may be associated with its role of anti-inflammation,anti-lipid peroxidation and down regulation of NF-κB protein level in lung tissue.
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<p><b>OBJECTIVE</b>To investigate the effects of initiative and passive perioperative function exercises on hidden blood loss (HBL).</p><p><b>METHODS</b>Two hundreds and thirty elderly patients with hip fractures aging from 67 to 87 years (average age of 73.6 years) who underwent total hip replacement were included. By the intensity and the manner of perioperative function exercises, patients were divided into four groups: little initiative function exercises group (group A, n=51), little initiative and passive function exercises group (group B, n=54), normal initiative function exercises group (group C, n=65), normal initiative and passive function exercises group (group D, n=60). The true total blood loss, HBL and their proportion on the original blood volume and total blood loss was calculated depending on height, weight, intra-operative blood loss, post-operative blood loss, pre- and post-operative hematocrit, and blood transfused. According to the proportion of mean HBL on total blood loss, patients were divided into low HBL group and high HBL group. The data were analyzed by t test.</p><p><b>RESULTS</b>The mean HBL was 517 ml, 41.9% of the total blood loss. Thereinto, the mean HBL was 695 ml in group A, 49.3% of the total blood loss, the prevalence of high HBL was 66.7% (34/51); the mean HBL was 625 ml in group B, 46.9% of the total blood loss, the prevalence of high HBL was 59.3% (32/54); the mean HBL was 446 ml in group C, 38.4% of the total blood loss, the prevalence of high HBL was 30.8% (20/65); the mean HBL was 346 ml in group D, 32.3% of the total blood loss, the prevalence of high HBL was 20.0% (12/60). Mean HBL, mean HBL/total blood loss, prevalence of high HBL were lower in group C than that in group A and group B (all P<0.05); and were lower in group D than that in group C (all P<0.05). The prevalence was 57.4% (132 cases) in low HBL group, and 42.6% (98 cases) in high HBL. The proportion of little initiative function exercises patients in high HBL group was obviously higher than that in low HBL group (P<0.05).</p><p><b>CONCLUSIONS</b>The intensity and the manner of perioperative function exercises are strongly associated with the HBL in elderly patients with total hip replacement. The initiative combined with the passive function exercises could be effectively prevent and reduce the incidence of high HBL.</p>
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Aged , Aged, 80 and over , Humans , Arthroplasty, Replacement, Hip , Blood Loss, Surgical , Exercise Therapy , Hip Fractures , General Surgery , Postoperative HemorrhageABSTRACT
BACKGROUND:The emergence of a large number of hidden blood loss during perioperative period of intertrochanteric fracture in the elderly not only increases the risk of perioperative period and complications, but also affects the postoperative recovery of joint function. At present, there is no relevant report about nutritional status and the hidden blood loss before surgery in and outside China. OBJECTIVE:To identify the effect of nutritional status on preoperative hidden blood loss in elderly patients with intertrochanteric fracture. METHODS:183 elderly patients with fresh and initial femoral intertrochanteric fracture were included. Laboratory serological examinations on admission and preoperation were completed. By mini nutritional assessment, patients were randomly divided into normal-nourishment group, malnourishment at risk group, and malnourishment group. The original blood volume and preoperative hidden blood loss were calculated depending on height, weight, hematocrit on admission and preoperation. According to the proportion of mean preoperative hidden blood loss on the original blood volume, patients were divided into low and high hidden blood loss groups. We compared preoperative hidden blood loss, and their proportion on the original blood volume and the preoperative incidence of high hidden blood loss, and analyzed the correlations between preoperative high hidden blood loss and preoperative nutritional status. RESULTS AND CONCLUSION:(1) The nutritional status of elderly intertrochanteric fracture patients measured by mini nutritional assessment score was that the number of patients was 48 cases (26%) in normal-nourishment group, 64 cases (35%) in the malnourishment at risk group, and 71 cases (39%) in the malnourishment group. There were no obvious differences in the preoperative complications between any two groups (P>0.05). (2) Thirty-eight cases affected high hidden blood loss. The mean preoperative hidden blood loss was 260.43 mL. The proportion of preoperative hidden blood loss to the original blood volume was 6%. (3) The preoperative hidden blood loss, their proportion on the original blood volume and the incidence of high hidden blood loss were significantly higher in the malnourishment at risk group and malnourishment group than in the normal-nourishment group. Paired comparison showed significant differences (P<0.05). (4) Results confirmed that preoperative hidden blood loss, their proportion on the original blood volume and the incidence of high hidden blood loss gradual y increased with deterioration of nutritional status. The nutritional status is an important factor influencing the occurrence of preoperative hidden blood loss, and can be used as an important index for judging the high hidden blood loss and prognosis in elderly patients with intertrochanteric fracture.
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Tension pneumopericardium is an uncommon complication of mechanical ventilaton. It may indeed be life-threatening for hemodynamic compromise and circulatory collapse. We present a case of tension pneumopericardium in a patient with a portable ventilator during intrahospital transport for computed tomography scan. Although timely rescue measures were performed, the patient died finally. We report the case to help us to be aware of and take precautions against this fatal condition during intrahospital patient transportation
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Objective:To screen the binding peptide against adenovirus type 7(Ad7) and evaluate the relevance with the ade-novirus receptor .Methods:Binding peptide against Ad 7 was screened by panning the phage display 12 peptides library .The antibody against the selected peptide was prepared and was used to study the binding to the membrane by immunofluorescence technique .Re-sults:Using Ad7 as the target protein , GTS09 peptide was selected from the phage display 12 peptides library by biopanning .GTS09-phage complex could significantly bind Ad 7, with the affinity constant up to 1.93 ×1010 L/mol;at the same time, immunofluorescence showed that antibody of GTS09 could specifically bind to membrane of 293 cell.Conclusion: Antibody against GTS09 peptide could specifically bind to membrane of 293 cell,which shows that the peptide may presumably have homology with the cell receptors of Ad 7.
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0.05).Spearman analysis revealed correlation between postoperative3 tibial plateau varus correction angle and functional recovery of knee joint(r=0.812,P