AIMTo investigate the effect of nimodipine (NM) on cerebral blood flow (CBF) in dogs following intranasal administration.

METHODSNM solution was administered intranasally, intravenously (i.v.), and orally to dogs and the change of CBF was determined by using electromagnetic blood flowmeter. MFLab experimental program was applied to monitor the experimental process and analyze data.

RESULTSCBF markedly increased after iv and intranasal application, while large variance was observed after oral dosing. CBF in dogs after three administrations increased by 26.4%, 28.0% and 8.5%, respectively, compared with that of baseline. Following intranasal administration, the onset of action was slightly slower than that after iv injection [(5 +/- 4) min vs (2.2 +/- 1.2) min], however the duration of improvement was the longest [ (25 +/- 17) min].

CONCLUSIONIntranasal delivery for NM can be a promising alternative to parenteral or oral administration.

Administration, Intranasal ; Administration, Oral ; Animals ; Blood Flow Velocity ; drug effects ; Cerebrovascular Circulation ; drug effects ; Dogs ; Female ; Injections, Intravenous ; Male ; Nimodipine ; administration & dosage ; pharmacology ; Vasodilator Agents ; administration & dosage ; pharmacology

To obtain the recombinant pZP3alpha protein for the study of the contraceptive vaccines, the DNA sequence (446-1423) encoding purified pZP3alpha was inserted into a vector--pPICZalphaA. The recombinant plasmid pPICZalphaA-pZP3alpha was linearized and then transformed into Pichia pastoris GS115 by electroporation. Engineering strains were attained by screening with zeocin and induced to produce rpZP3alpha in high-density fermentation. Then rpZP3alpha was purified by Cu2+ metal affinity column chromatography from the separated and concentrated fermentative supernatants. The purified rpZP3alpha was identified by SDS-PAGE and Western blot, and the quantity, purity and rate of recovery of the rpZP3alpha were analyzed by Quantity One software. One male rabbit was immunized with the Cu-NTA-purified rpZP3alpha. The antibody responses against rpZP3alpha and porcine ZP were detected by ELISA and the indirect immunofluorescence. Engineering strains expressing rpZP3alpha in secretion were constructed. A 46kD component named rpZP3alpha which can react with anti-pZP3 antibody was purified from fermentative supernatants of engineering strains and the average yield of purified rpZP3alpha obtained from fermentative supernatants was 8mg/L. The purity and the rate of recovery were up to 92% and 63% respectively. The anti-rpZP3alpha antiserum was prepared by immunization of a male rabbit with purified rpZP3alpha. This anti-rpZP3alpha antiserum could react with rpZP3alpha and purified pZP3 in ELISA and bind to porcine zona pellucida which produced bright green fluorescence in the indirect immunofluorescence. The rpZP3alpha (46kD) protein could be successfully expressed in the Pichia pastoris expression system. And this protein retained the immunogenic activity of natural pZP3.
Animals ; Egg Proteins ; genetics ; metabolism ; Electroporation ; Fermentation ; Immunization ; Male ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Rabbits ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; secretion ; Swine ; Zona Pellucida Glycoproteins

Objective:To explore the predictive value of quantitative electroencephalogram (qEEG) in the poor outcome of children with non-traumatic disturbance of consciousness (DoC) in the pediatric intensive care unit (PICU).Methods:A prospective study was conducted. From January 2019 to May 2019, a total of 62 patients aged from 1 month to 11 years with non-traumatic DoC in the PICU of the First Affiliated Hospital of Bengbu Medical College were enrolled. Bedside monitoring with NicoletOne monitor was performed within 24 hours after admission, and qEEG parameters, including amplitude-integrated electroencephalogram (aEEG), relative alpha variability (RAV), relative band power (RBP), and spectral entropy (SE) were recorded. The state of consciousness was assessed with modified pediatric Glasgow coma scale (MPGCS) before monitoring. According to the pediatric cerebral performance category score at 1 year after discharge, the enrolled subjects were divided into good and poor outcome groups. The association between these variables and the poor outcome was analyzed by univariate and multivariate logistic regression analysis, and the predictive performance was analyzed by receiver operator characteristic (ROC) curve.Results:There were 39 males and 23 females, with the age of 12.0 (5.8, 24.0) months. Fifty patients (81%) were in the good outcome group and 12 patients (19%) in the poor outcome group. The univariate Logistic regression analysis showed that age ( OR=1.037, 95% CI 1.001-1.074, P=0.041), severe abnormal aEEG ( OR=128.000, 95% CI 10.274-1 594.656, P<0.01), RAV ( OR=0.877, 95% CI 0.810-0.949, P=0.001), SE ( OR=0.892, 95% CI 0.814-0.978, P=0.015), and MPGCS score ( OR=0.511, 95% CI 0.349-0.747, P=0.001) were significantly associated with the poor outcome. However, the multivariate Logistic regression analysis showed that only severe abnormal aEEG ( OR=315.692, 95% CI 6.091-16 362.298, P=0.004) and RAV ( OR=0.808, 95% CI 0.664-0.983, P=0.033) were significantly associated with the poor outcome. The area under the curve (AUC) of the aEEG and RAV in predicting the poor outcome were 0.848 (95% CI 0.735-0.927, P<0.01) and 0.847 (95% CI 0.733-0.926, P<0.01), respectively. The optimal cut-off value was severe abnormal for the aEEG and 38% for the RAV, with sensitivity of 67% and 83%, specificity of 98% and 84%, positive predictive value of 89% and 55%, negative predictive value of 92% and 95%, and Youden index of 0.647 and 0.673, respectively. The AUC of the novel combined index of aEEG and RAV for predicting the poor outcome was 0.974 (95% CI 0.898-0.998, P<0.01). Conclusions:The aEEG and RAV are reliable predictors for the poor outcome of children with non-traumatic DoC, and the novel combined index of aEEG and RAV can improve the predictive performance. The qEEG can be used as a routine method for outcome assessment due to its good objectivity.

OBJECTIVETo investigate the effect of docosahexaenoic acid (DHA) on invasiveness of aflatoxin B1 (AFB1)-induced hepatocellular carcinoma cells in vitro.

METHODSHepG2.2.15 cells were exposed to different concentrations of AFB1 and DHA plus AFB1. The cell migration and invasion were assessed using wound-healing and Transwell assay, and flow cytometry was used to analyze the cell cycle changes. The ultrastructural changes of the cells were observed by transmission electron microscopy.

RESULTSCompared with the control group, the cells exposed to2 µmol/L AFB1 showed obviously enhanced migration and invasion with decreased cell ratio in G1/G1 phase and increased cell ratio in G2/M phase but no changes in S phase cells; transmission electron microscopy revealed the presence of multiple nucleoli and significantly increased mitochondria and Golgi apparatus in the exposed cells. Compared with AFB1-exposed cells, the cells treated with DHA and AFB1 showed decreased migration and invasion abilities, and the G1/G1 phase cells increased and G2/M phase cells decreased significantly; ultrastructurally, the cells contained single nucleoli with decreased mitochondria and vacuolization occurred in the cytoplasm.

CONCLUSIONDHA can significantly inhibit AFB1-induced enhancement of cell migration and invasion in hepatocellular carcinoma cells in vitro.

Aflatoxin B1 ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Cycle ; Cell Movement ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Golgi Apparatus ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; Mitochondria ; Neoplasm Invasiveness

OBJECTIVETo study the feasibility of inducing the differentiation of rat adipose-derived stem cells (ADSCs) into smooth-muscle-like cells in monolayer culture in vitro.

METHODSADSCs were obtained from the adipose of the inguinal region of the SD rat. A growth curve of the ADSCs was drawn. The fourth passage ADSCs were induced to differentiate into adipocytes with adipogenic inducing fluid and determined by Oil Red O staining, into osteoblasts with osteogenic inducing fluid and determined by Von Kossa staining, as well as into smooth-muscle-like cells with inducing fluid containing beta-mercaptoethanol, and the expression of alpha-smooth muscle actin (alpha-SMA) was detected by the immunohistochemical method.

RESULTSThe ADSCs presented spindle and polygon shapes and proliferated rapidly in vitro. The growth curve showed the ADSCs in the logarithmic growth phase two days after subculture. The fourth passage ADSCs exhibited red stained lipid droplets characteristic of adipocytes in the cytoplasm on Oil Red O staining after induction with adipogenic inducing fluid, and calcium nodes on Von Kossa staining after induction with osteogenic inducing fluid. The immunohistochemical results of the ADSCs induced into smooth-muscle-like cells showed that the positive rate of alpha-SMA in the beta-mercaptoethanol induced cells was (29.80 +/- 6.89)%, significantly higher than in the uninduced ones ([2.89 +/- 1.24]%, P < 0.01).

CONCLUSIONADSCs displayed obvious characteristics of smooth muscle cells after induction, and could be a new source of cells in the tissue engineering studies of smooth muscle related diseases.

Adipocytes ; cytology ; Animals ; Cell Culture Techniques ; Cell Differentiation ; Male ; Myocytes, Smooth Muscle ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the safety of living related donors in short term after transplantation.

METHODSTwo hundred and fifty-one cases of living related donor kidney transplantation from May 2000 to July 2007 were analysed retrospectively. There were 117 male and 134 female aged from 22 to 72 years old, with a mean of 46.6 years old. The indexes were compared including serum creatinine (SCr), creatinine clearance (CCr), glomerular filtration rate (GFR) and quality of life before and after donation. Surgical complications were followed-up.

RESULTSDonors' SCr was (75.9 +/- 17.2) micromol/L before donation, (107.4 +/- 21.2) micromol/L on 7 d after donation, (130.4 +/- 58.2) micromol/L at the 1(st) month and (116.1 +/- 24.1) micromol/L at the 3(rd) month. There were significant difference between any 2 time points (P < 0.01). CCr was (94.4 +/- 17.5) ml/min before donation and (63.5 +/- 17.8) ml/min on 10 d after donation (P < 0.01). In 62 donors, total GFR was (82.4 +/- 21.8) ml/min before donation. On 10 d after donation, GFR of remaining kidney was (57.4 +/- 14.1) ml/min which was 34.7% higher than GFR of this kidney before donation (42.6 +/- 11.8) ml/min. There was no significant difference in quality of life before living related donors and non-donor populations (P = 0.116). Surgical complications included splenic rupture in 1 case, descending colon rupture in 1 case and wound infection in 5 cases.

CONCLUSIONLiving donor kidney transplantation is safe for donors, although part of indexes would vary within normal range during the early time after donation.

Adult ; Aged ; Female ; Humans ; Kidney Transplantation ; adverse effects ; Living Donors ; Male ; Middle Aged ; Nephrectomy ; adverse effects ; Postoperative Period ; Retrospective Studies ; Safety ; Young Adult

BACKGROUND: Proximal femoral nail antirotation (PFNA) is the most commonly used internal fixation for femoral intertrochanteric fracture. However, a large amount of hidden blood loss makes negative effect on the functional recovery postoperatively. OBJECTIVE: To compare the clinical efficacy of the modified incision and standard incision in PFNA for femoral intertrochanteric fracture. METHODS: Sixty-nine patients with femoral intertrochanteric fractures were analyzed retrospectively, and were then divided into two groups, followed by treated with modified incision of PFNA (group A) or standard incision of PFNA (group B). The postoperative biomechanical stability, fracture healing time, complications, Harris scores at the last follow-up, proximal incision length, operation time, and blood loss were compared between two groups. RESULTS AND CONCLUSION: (1) All the patients were followed up for more than 12 months. The fracture healing time, Harris scores at the last follow-up and incidence of postoperative complications did not differ significantly between two groups. (2) The intraoperative blood loss, total blood loss and total blood transfusion in the group A were significantly less than those in the group B (P < 0.05). (3) The proximal incision length and operation time in the group A were significantly shorter than those in the group B. (4) These results manifest that in the treatment of femoral intertrochanteric fracture, both methods exhibit good biomechanical properties and stability. Notably, compared with the standard incision method, the modified incision method has less trauma, shorter operation time, less blood loss and blood transfusion.

In order to explore the effect and its mechanism of paeoniflorin on PD-L1, a PD-L1 high expression cell model was established in interferon gamma(IFN-γ)-induced HepG2 cells. The cytotoxicity of paeoniflorin was detected by MTT assay. Flow cytometry, ELISA and RT-PCR were performed to detect protein and mRNA levels of PD-L1 regulated by paeoniflorin. In HepG2 cells and Jurkat T cell co-culture system, the expression of IL-2 was detected by ELISA. Besides, T cell proliferation was evaluated by CCK-8 method, and the protein expression levels of PD-L1, JAK and STAT3 after drug treatment were determined by Western blot. These results indicated that paeoniflorin could significantly down-regulate the levels of PD-L1 protein and mRNA. In addition, it increased the number of T cells and the concentration of IL-2 in the co-culture system. Furthermore, paeoniflorin could significantly inhibit the protein expression of JAK and STAT3. Au the above experimental data indicated that paeoniflorin could down-regulate the expression of PD-L1, and its mechanism might be related to the JAK/STAT3 pathway.

Objective: To isolate and identify Prevotella nigrescens (P.nigrescens) in gingival crevicular fluid of patients with chronic periodontitis and to analyze its tumor-promoting role in esophageal squamous cell carcinoma (ESCC). Methods: Samples of gingival crevicular fluid were collected from patients with chronic periodontitis and cultured on GAM agar medium under anaerobic conditions. Black colonies on GAM agar plates were picked for subculture, and then the bacteria were isolated and purified. Bacterial identification was conducted using Gram staining and 16S rDNA sequencing analysis. The isolated bacterial species was used to infect ESCC NE6-T cells and the changes in biological characteristics of the infected cells were evaluated. Results: Multiple bacterial colonies were observed after anaerobic culturing of gingival crevicular fluid samples for 120 h on GAM agar plates. A single bacterial colony with pure black and smooth appearance was obtained from grey black bacterial colonies with streak method. It was a Gram-negative bacterium with bead-like shape. It showed 99.78% 16S rDNA sequence identity with P. nigrescens F0103 and was named P. nigrescens LY01. P. nigrescens LY01 infection promoted the in vitro proliferation, migration and invasion of NE6-T cells and the growth of subcutaneous xenograft in nude mice. In addition, it could also induce the upregulation of Ki67 and activation of p-STAT3. Conclusions P. nigrescens inhabiting in gingival sulcus might promote the progression of ESCC in human with chronic periodontitis.

Objective:To investigate the relationship between tube voltage kV value for head and neck CT angiography and body weight.Methods:A total of 120 patients with suspected vascular disease of the head and neck who underwent CT angiography of the head and neck in Beihai People's Hospital from January 2020 to May 2022 were included in this study. Patients were divided into three groups according to different tube voltages: group A (tube voltage 120 kV, n = 45), group B (tube voltage 100 kV, n = 45) and group C (tube voltage 80 kV, n = 30). Patients in group A were divided into group A1 (< 70 kg, n = 15), group A2 (70-85 kg, n = 15) and group A3 (> 85 kg, n = 15) according to different body weights. Patients in group B were divided into group B1 (< 70 kg, n = 15), group B2 (70-85 kg, n = 15) and group B3 (> 85 kg, n = 15) according to different body weights. Patients in group C were divided into group C1 (< 70 kg, n = 15) and group C2 (70-85 kg, n = 15) according to different body weights. Group C3 was not used. The contrast medium used was Loversol. The CT value, image noise, signal-to-noise ratio, contrast to noise ratio, and effective radiation dose of arterial vessels in each group were measured. The images were subjectively evaluated by two physicians who had senior professional titles using a 5-point rating scale. Results:Subjective score of image quality was all ≥ 3 grade in each group. There was no significant difference in image quality rating between groups A1 and A2 and groups B1, B2, and C1. There was a remarkable difference in image quality rating between groups A3, B3, and C2 and the other groups. There was a significant difference in the CT value of blood vessels at four different levels between groups A1, B1 and C1 ( F = 76.82, 64.62, 98.79, 71.85, all P < 0.001). There was a significant difference in CT value of blood vessels at four different levels between groups A2, B2 and C2 ( F = 159.82, 112.33, 108.22, 135.18, all P < 0.001). There was a significant difference in CT value of blood vessels at four different levels between groups A3 and B3 ( t = 4.40, 4.27, 3.91, 3.59, all P < 0.05). In groups B3 and C2, the image noise was remarkably increased, signal to noise ratio and contrast to noise ratio were remarkably decreased compared with those in the other groups. The effective radiation dose of arterial vessels in group B1 was 47% lower than that in group A1 and the effective radiation dose of arterial vessels in group C1 was 73% lower than that in group A1 ( F = 116.18, P < 0.001). The effective radiation dose of arterial vessels in group B2 was 49% lower than that in group A2, and the effective radiation dose of arterial vessels in group C2 was 66% lower than that in group A2 ( H = 35.40, P < 0.001). The effective radiation dose in group B3 was 35% lower than that in group A3 ( t = 3.59, P < 0.05). Conclusion:In CT angiography of the head and neck, the selection of tube voltage kV value is related to body weight. Tube voltage 80 kV is suitable for patients with a body weight < 70 kg, tube voltage 100 kV for patients with a body weight of 70-85 kg, and tube voltage 120 kV for patients with a body weight > 85 kg. These tube voltages can decrease effective radiation dose and ensure image quality, meeting the requirement for clinical diagnosis.