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Objective:To prepare rabbit polyclonal antibodies against respiratory syncytial virus (RSV) N protein and use them as the detection antibodies to establish a N-ELISA-based method for rapid detection of neutralizing antibodies.Methods:A plasmid of pET30a-N for the expression of RSV N protein was constructed. After purification, the protein was immunized into New Zealand rabbits to prepare polyclonal antibodies, which were used as the detection antibodies. Positive serum samples were diluted and used to neutralize RSV (100 TCID 50/well). Hep-2 cells were inoculated and cultured, and then the cells were fixed with 80% acetone. ELISA was performed to detect RSV N protein in infected cells. When the absorbance value of a well was below the cut-off value, it was regarded as the positive well in the neutralization test. The highest dilution of a positive well serum was the neutralizing antibody titer. After optimizting the antibody dilution, detection time, cell density and the duration of neutralization, the method for neutralizing antibody detection was established based on N-ELISA. The established method was verified by analyzing the influences of different cell generations and edge effects, and calculating the accuracy, repeatability and precision. The correlation between the established method and microneutralization method was analyzed by detecting human RSV IgG-positive serum. Results:The plasmid pET30a-N was successfully constructed, and the expressed N protein showed high purity and good specificity. After the third immunization, the antibody titer in rabbit serum was 1∶51 200, and the antibodies could specifically bind to RSV. The prepared rabbit anti-RSV N polyclonal antibodies had a titer of 1∶51 200, and showed good specificity. The neutralizing antibodies could be detected on day 4 with the established method, and the duration of neutralization was shortened to 30 min. Cell generations and the position of wells in the 96-well plate (edge well and non-edge well) had no significant effect on the method, and the repeatability, precision and accuracy of the method were good. In the detection of 64 RSV IgG-positive human serum samples by the established method and microneutralization method, the correlation coefficient was 0.929 6, indicating a good positive correlation between the two methods.Conclusions:A N-ELISA-based method for rapid neutralizing antibody detection is successfully established, which can be used to evaluate the serum antibody level after RSV vaccination.
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Respiratory syncytial virus (RSV) is one of the major pathogens of acute respiratory infections, becoming a huge global burden. Virus-receptor interactions play a key role in the pathogenesis of RSV infection. The distribution of receptors influences the cellular and the tissue tropism of RSV as well as the viral replication and proliferation in the host. However, the RSV receptors are currently unknown, which is one of the reasons that hinders the development of RSV vaccines and therapeutic drugs. In this study, the existing RSV receptors are reviewed with the hope to provide ideas for the research of RSV vaccines and therapeutic drugs.
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Objective To investigate the conversion rate, organ procurement rate and influencing factors of organ donation in a single center from Fujian province. Methods Baseline data of 182 potential organ donors of Fujian Medical University Union Hospital from November 2018 to June 2021 were retrospectively analyzed. The conversion rate of organ donation, baseline data of successful organ donors and the causes of failure of organ donors were identified. The organ procurement rate and the influencing factors of the number of organ donations were analyzed. Results Among 182 potential organ donors, 46 cases were successful organ donors with a conversion rate of 25.3%. In addition, 136 cases failed to donate organ. The main causes included disagreement from family members (58.1%), insufficient evaluation time (24.3%) and ineligible for donation criteria (17.6%). A total of 212 large organs and tissues were donated by 46 organ donors, including 88 kidneys, 42 livers, 15 lungs, 19 hearts and 48 corneas, with 4.6 large organs and tissues, and 3.6 large organs for each donor. Age, sex, native place, organ donation area and blood type were the influencing factors of the number of large organ donations. Organ donation area was the influencing factor of the number of tissue donations. Conclusions The conversion rate of organ donation is relatively low in a single center from Fujian province. Disagreement from family members is the main cause. Suitable potential organ donors should be selected for organ donation to improve the conversion rate and organ procurement rate of organ donation.
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Objective:To establish a predictive model of moderate to severe pain in patients with hepatocellular carcinoma (HCC) undergoing transcatheter arterial chemoembolization (TACE).Methods:264 patients with HCC who underwent TACE operation in Southern Medical University from January 2017 to April 2018 were selected as the modeling set. The pain was assessed by numeric rating scales. The patients were divided into pain group ( n=96) and non-pain group ( n=168) according to whether moderate to severe pain occurred within 24 hours after the operation. Binary Logistic regression analysis were performed for variables that were statistically significant in the univariate analyses. The predictive nomogram was constructed and the internal validation was performed. In addition, 87 patients with HCC who underwent TACE operation from January 2020 to June 2020 were selected as the validation set for external validation. Results:In the modeling set, 96 patients (36.36%) had moderate to severe pain within 24 hours after TACE operation in 264 patients with HCC, and the dosage of morphine intramuscularly injected within 24 hours was 1015 mg, with an average of 10.57 mg per patient. Multivariate Logistic regression analysis showed that preoperative pain, the distance between the tumor and capsule ≤2 cm, high prothrombin activity, dosage of lipiodol>10 ml, and several thromboembolic tumors were independent risk factors for moderate to severe pain after TACE ( P<0.05). Age>50 was the protective factor of moderate to severe pain after TACE ( P<0.05). The area under ROC curve was 0.799 (95% CI: 0.745-0.853) in the modeling set. The area under Roc curve for internal validation and external validation were 0.780 and 0.788, respectively. The calibration curves showed satisfactory agreements between the model predicted probability and the actually observed probability. Conclusion:The predictive model of moderate to severe pain after TACE was established in this study has good differentiation and accuracy, it has certain guiding significance for predicting the high-risk group of moderate to severe pain after TACE operation and formulating the targeted prevention strategy.
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Objective:To understand the attitudes of obstetricians and midwives towards the development of nurse practitioner in China.Methods:The phenomenological method in qualitative research was used to conduct semi-structured interviews with 24 medical staff in the third-A general hospitals and women and children's hospitals in Guangzhou and Dongguan City. The interview data were analyzed by Colaizzi's 7-step analysis method.Results:Four themes were extracted, namely, unanimous approval attitude, imperfect midwifery discipline and management system, insufficient independent practice capacity, low support and recognition in midwifery area.Conclusions:Obstetricians and midwives hold a positive attitude towards the development of midwifery nurse practitioner. At the same time,its development is restricted in many aspects. They can gradually develop midwife′s legislation, improve the management system, try some prescription rights, and increase the education and talents of midwifery subjects. Cultivation and promotion of social recognition will help gradually train midwifery nurse practitioner in China.
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Objective To investigate the relationship between electrical stimulus intensity and postopera-tive complications of puncture trigeminal ganglion radiofrequency thermocoagulation (PTGRT). Methods 86 pa-tients with trigeminal neuralgia (TN) on the second and the third branch underwent trigeminal ganglion radiofre-quency thermocoagulation in Xuanwu Hospital , Capital Medical University from June 2013 to December 2015 enrolled in this study. Electrical stimulus intensity in surgery and postoperative pain VAS score , numbness score, masseter muscles score and complications were recorded at 0, 24 and 48 hours post-surgery. Correlations among electrical stimulus intensity in surgery and postoperative pain VAS score,numbness score, masseter mus-cles score were analyzed. Result The 86 patients had different degrees of facial numbness , some patients had different degrees of masseter weakness , and 2 patients had mild sialorrhea in two days after PTGRT without serve complications. Eighty-three patients (96.51%) had no pain, and VAS score wass 0 at 0, 24 and 48 hours post-PTGRT. Electrical stimulus intensity in surgery had apparent negative correlation with numbness score at 0 , 24 and 48 hours popst-PTGRT (P < 0.01), with negative correlation with masseter muscles decreasing at 48 hours post-PTGRT (P < 0.05). Conclusion Suitable temperature and time in PTGRT were needed to be chose ac-cording to the electrical stimulus intensity.
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Objective To observe the dynamic changes of brain edema,tumor necrosis factor-α (TNF-α),aquaporin-4 (AQP-4) in focal cerebral ischemia rats and investigate the effect of atorvastatin on the above-mentioned factors.Methods One hundred and eight male SD rats were randomly divided into sham-operated group,model group and treatment group (n=36);furthermore,each group was divided into three subgroups:1 d group,3 d group and 7 d group (n=12).Focal cerebral ischemia models in model group and treatment group were induced by left middle cerebral artery occlusion,while rats in the sham-operated group were only separated from blood vessels with no ligation and no inserted line.After succeeded models,the treatment group received gavage administration of atorvastatin (4 mg/ [kg·d]),others were given the same dosage of normal saline at the same time.The brain edema was calculated by measuring brain water content via dry-wet method,and the mRNA expressions of TNF-α and AQP-4 were detected by real time-PCR;Western blotting was employed to detect the AQP-4 protein expression.Results At the same observation time,as compared with those in the sham-operated group,the brain water content,TNF-α mRNA expression and A QP-4 mRNA and protein expressions in the model group and treatment group markedly increased (P＜0.05);as compared with the model group,treatment group had statistically decreased brain water content,TNF-α mRNA expression and AQP-4 mRNA and protein expressions (P＜0.05).Conclusion The brain edema of focal cerebral ischemia rats may be positively correlated to AQP-4 mRNA and protein expressions induced by TNF-α;atorvastatin probably down-regulates AQP-4 mRNA and protein cxpressions by inhibiting TNF-α to alleviate brain edema and protect brain tissues.
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Objective To investigate the effect ofatorvastatin on angiogenesis of brain tissues in focal cerebral ischemia rats,and explore the corresponding mechanism.Methods Ninety male SD rats were randomly divided into sham-operated group,model group and treatment group (n=30);and then,each group was divided into three subgroups:1 d group,3 d group and 7 d group (n=10).The focal cerebral ischemia models were induced by middle cerebral artery occlusion (MCAO);3 h after MCAO,rats in the treatment group received a gavage of atorvastatin 4 mg/(kg.d),and others were given the same amount of normal saline at corresponding time.The nerve function defects were estimated at 3 h after MCAO and before being killed;the protein expressions of vascular endothelial cell markers CD105 and glucose-regulated protein 78 (GRP78) were analyzed by immunohistochemistry;the vascular endothelial growth factor (VEGF) mRNA expression was analyzed by real time-PCR;the caspase-12 protein expression was analyzed by Western blotting.Results (1) Nerve function defects scores:there was no significantly statistical difference between model group and treatment group at 3 h after MCAO (P＞0.05),but statistical differences at different time points before being killed were noted (P＜0.05).(2) Microvessel density (MVD):at all time points,that of model group was increased as compared with that of sham-operated group,that of treatment group was increased as compared with that of model group,and the differences were all statistically significant (P＜0.05).(3)The VEGF rmRNA expression gradually increased over time;at all time points,the VEGF mRNA expression of model group was significantly higher than that in the sham-operated group,and that of treatment group was significantly higher than that in the model group (P＜0.05).(4)The GRP78 and caspase-12 expressions were gradually decreased over time;at all time points,the GRP78/BiP and caspase-12 expressions in the model group were significantly higher than those in the sham-operated group;those in the treatment group were statistically lower than those in the model group,but obviously higher than those in the sham-operated group (P＜0.05).Conclusion The angiogenesis of brain tissues in MCAO rats is obvious,and atorvastatin can enhance this effect;the mechanism may be that atorvastatin can weaken the endoplasmic reticulum stress,and then,reduce the apoptosis of endothelial cells.
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Objective To investigate the association of prehepatic fat thickness with insulin resistance in women with polycystic ovarian syndrome (PCOS).Methods Thirty-five PCOS patients and 39 controls were randomly enrolled in the study.Prehepatic fat thickness was detected by ultrasound,and fasting glucose (FBG),fasting insulin (FINS),and serum lipids were detected.The homeostasis model assessment of insulin resistance (HOMA-IR) was used to calculate insulin resistance.Results Prehepatic fat thickness was significantly higher in PCOS patients than healthy controls,and prehepatic fat thickness were significantly correlated with HOMA-IR (P ＜ 0.05).The receiver operating characteristic (ROC) curve demonstrated that prehepatic fat thickness had higher sensitivity and specificity in diagnosing PCOS with insulin resistance.Conclusions This study demonstrates that prehepatic fat thickness significantly correlates with insulin resistance and can be used as the markers of insulin resistance in PCOS patients.
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AIM:To observe protective effect of L-glutamine on myocardial cell injury induced by endotoxin. METHODS:22 male SD rats,weighting(250?30)g,were randomly divided into three groups:control group( n= 7),endotoxin group( n= 7),glutamine/endotoxin group( n= 8). Heart were isolated from rats and perfused on Langendorff apparatus with Krebs- Henseleit(K-H)buffer(Saturation 95%O 2+5%CO 2)at a constant pressure(8.33 kPa)and temperature(37℃). The activity of superoxide dismutase (SOD)and malondialdehyde (MDA)content of efflux from coronary artery, monophasic action potential(MAP)and contractile force were measured at certain timepoints (0 min,20 min,50 min,80 min). RESULTS: Monophasic action potential and contractile force were markedly improved in Gln/ET group. The activity of SOD and MDA level in Gln/ET group restored closely to that in control group. CONCLUSION:L-gluta mine protects myocardial cells from injure induced by endotoxin.
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AIM: To detect the role of fibrinogen activity (Fa) in the progress of coronary artery disease (CHD).METHODS:Fa was measured with hemorheology methods in patients with CHD stable phase (n=30) and angina pectoris (n=27).RESULTS: (1) Levels of plasmatic fibrinogen and plasmatic viscosity in patients with CHD were higher than that of control group (P