Objective To prepare rabbit polyclonal antibodies against pertussis toxin(PT) and develop a double antibody sandwich ELISA for quantitative determination of PT antigen,identify and apply the method.Methods The rabbit polyclonal antibody against PT was prepared by immunizing Chinchilla rabbit with PT using traditional method.The reaction conditions of ELISA system were optimized,the double antibody sandwich ELISA method for quantitative determi-nation of PT was developed,and the specificity,linearity,accuracy,precision and sensitivity were verified.The developed method was used to detect PT antigen content in fimbriae proteins(FIM) stock solution of samples during detoxification and other purification process of pertussis antigen.Results The working condition of double antibody sandwich ELISA for detection of PT antigen content was the coating concentration of PT rabbit polyclonal antibody of 1 μg/mL,and the enzyme-labeled antibody dilution of 1:8 000.This detection system showed specific reaction with PT purified protein,but had no cross reaction with filamentous hemagglutinin,diphtheria toxoid and tetanus toxoid;the linear detection range of the developed double antibody sandwich ELISA was within 25—400 ng/mL;the recovery rates of PT at high,moderate and low concentrations were 103.27%,91.48% and 103.52%,respectively;both the intra-and inter-coefficients of variation(CVs)were less than 10%;the sensitivity of the method was 20.719 ng/mL,and the detection limit was 41.438 ng/mL.Thirty-five batches of samples were detected under five different detoxification process conditions and at different sampling time points,and the changes of antigen content were all consistent with the trend of detoxification reaction.Conclusion The PT rabbit polyclonal antibody was successfully prepared,and a double antibody sandwich ELISA with high precision and accuracy was developed for the quantitative determination of PT antigen content,which can be used for the antigen content detection of chemically detoxified samples in the production process of component DPT vaccines.

The advantages of saliva-based biomarkers detection in cancer were accurate,simple and noninvasive.Currently,biomarkers validated for saliva detection include protein,mRNA,miRNA and DNA,using PAGE,microarray and sequencing,respectively.Analysis of literatures shows;that saliva biopsy plays an important role in cancer diagnosis,clinical treatment and prognosis.Due to the design of experiments reported werediversification,a large number of validations are necessary to standardize saliva collecting,processing,testing methods and quality assurance.

Objective: To investigate the mechanism of scorpion venom active peptides(SVAPs) on platelet aggregation. Methods: Platelet aggregation in vivo and vitro was determined by turbidimetry. Carotid thrombosis model was induced by electrostimulation. The determination of 6 keto PGF 1 and TXB 2 were performed by radioimmunoassay. Results: SVAPs 0.125,0.25,0.5mg?ml -1 significantly inhibited the rabbit platelet aggregation triggered by thrombase 0.03u.ml -1 , ADP 10u.ml -1 in vitro( P

Objective: To explore the gender related differences of clinical symptoms and triggering factors in patients with acute myocardial infarction (AMI) in China.
Methods: A population of 14 854 AMI patients with CAMI registration from 2013-01 to 2014-03 were studied, which included 10999 (74.0%) male and 3855 (26.0%) female. The gender related differences of clinical symptoms and triggering factors were analyzed in the population.
Results: The chest pain and severe sweating were the most common symptoms of AMI patient, there were 66.4%of patients with persistent chest pain and 63.7%of patients with severe sweating. Male patients were more with chest pain (67.8%vs 62.4%) and severe sweating (65.8%vs 58.0%) than female, while female patients were more with radiating pain (36.0%vs 31.0%) and nausea/vomiting (35.6%vs 25.0%) than male, all P<0.05. There were 19.4%of patients having clear cause of AMI, and physical stress was the most frequent trigger (48.5%) for AMI. Male patients usually had clear cause than female (20.9%vs 15.1%), especially because of physical stress (49.5%vs 44.5%) and excessive recent unhealthy lifestyles (15.4%vs 8.8%), all P<0.05.
Conclusion: Chest pain and severe sweating were the most common clinical symptoms for AMI patients in China, about 1/5 of them had triggering factors and it was more in male patients.

Objective To investigate the clinical effects of two methods of compressive coverage with aseptic gauze dressing and alginate dressing on haemostasis in placement of peripherally inserted central catheter(PICC)so as to provide a new dressing method for clinical application.Methods A total of 416 breast cancer patients undergoing PICC were randomly divided into two groups,with 208 per group.In the control group,the puncture sites for PICC were dressed with aseptic gauze immediately after the placement of PICC,with a daily aseptic gauze dressing change once per 24 hours.In the trial group,the puncture sites were dressed with alginate dressing immediately after the placement of PICC,with a weekly dressing change at the 7th day after PICC placement,while no local bleeding on the puncture site.The incidence,within 24 hours,of bleeding,degree of comfort,frequency and cost of dressing change as well as the infection rate within 7 days after the PICC placement were monitored and compared between the groups.Results A total of 205 patients in the control group and 208 in the observation group had completed the study.In the trial group,the incidence of bleeding at puncture sites within 24 hours after placement of PICC was significantly lower than that of the control group(27.88%vs.38.05%,χ2=4.829,P=0.028).The frequency of dressing change within 24 hours in the observation group was significantly less than that of the control group(Z=205.235,P<0.001).The cost of dressing change in the observed groups was also significantly less than that of the control group(56.94 vs.10.20 yuan in average,Z=-8.990,P<0.001).The incidence rates of local pain and itche reported by the patients in the observation group were significantly lower than those in the control group(Z=-12.079,P<0.001;Z=-12.194,P<0.001).No infection at the puncture site was observed in both groups.Conclusions Application of alginate dressing immediately after PICC placement in breast cancer patients can extend the time for initial dressing change up to 7 days in the patient without bleeding at the puncture site within 24 hours after the placement of PICC.An alginate dressing can reduce the incidence of bleeding at the puncture site,increase the comfort of patients,lower the frequency and cost of dressing change hence the nursing workload.

Objective:The activation of astrocytes is an important process in the formation of chronic pain.This study aims to observe the activation of A1 reactive astrocytes in the medullary dorsal horn in the rat model of trigeminal neuralgia,and to explore the mechanism of central sensitization caused by A1 reactive astrocyte. Methods:The adult male rats were randomly divided into a sham group and a chronic constriction injury of infraorbital nerve(ION-CCI)group.The facial mechanical pain threshold and thermal withdrawal latency were measured before the operation and on the 1st,3rd,7th,10th,and 14th day after the operation.After pain behavior observation,the expression of glial fibrillary acidic protein(GFAP)in the medullary dorsal horn was observed by immunohistochemistry and immunofluorescence colocalization of GFAP,complement 3(C3)/S100A10,and 4',6-diamidino-2-phenylindole(DAPI)was analyzed.Primary astrocytes were cultured and randomly divided into a naive group and a DHK group.The DHK group was treated with 1 mmol/L of astrocyte activation inhibitor dihydrokainic acid(DHK).Fura-2/AM was used to stain the astrocytes and the calcium wave of the 2 groups under the stimulation of high potassium was recorded and compared.The expression of C3 was detected by Western blotting. Results:The facial mechanical pain threshold and thermal withdrawal latency of the ION-CCI group were significantly lower than those of the sham group(both P<0.05).There were a large number of GFAP positive astrocytes in the medullary dorsal horn of the ION-CCI group.The fluorescence intensity of GFAP in the ION-CCI group was higher than that in the sham group(P<0.05).GFAP and C3/S100A10 were co-expressed in astrocytes.Compared with the sham group,the fluorescence intensity of C3 and the protein expression of C3 in the ION-CCI group were increased(both P<0.05).The expression of C3 in ION-CCI group was significantly increased(P<0.05).Compared with the naive group,the C3 protein expression was significantly decreased in the DHK group(P<0.05).The intensity of calcium fluorescence was increased after high potassium stimulation in both groups.Furthermore,the peak and increase amplitude of calcium fluorescence in the naive group were much higher than those in the DHK group(both P<0.05). Conclusion:A1 reactive astrocytes in the medullary dorsal horn of trigeminal neuralgia model rats are increased significantly,which may participate in central sensitization of trigeminal neuralgia by impacting astrocyte calcium wave.

OBJECTIVE: To investigate the therapeutic efficacy of coblation-assisting uvulopalatopharyngoplasty (CAUPPP) combining with coblation-channeling of the tongue (CCT) for patients with severe obstructive sleep apnea-hypopnea syndrome (OSAHS). METHOD: A retrospective analysis was made on patients with severe OSAHS treated by CAUPPP combining with CCT. The adult with severe OSAHS were involved in the study and the apnea-hypopnea index (AHI) were beyond 40/h and lowest arterial oxygen saturation (LSaO2) were under or equal to 80%. Surgical approach: The UPPP was performed to decrease the size of soft palate with coblation and coblation-channeling in the soft palate and decrease the size of tongue by CCT. All patients were followed up for 6 to 12 months and underwent polysomnography (PSG). RESULT: Subjective symptoms of patients improved more significantly than that of preoperation. The function of soft palate is normal without significant nasopharyngeal regurgitation. Compared with the preoperative data, AHI values were significantly decreased (P<0.01) and the lowest oxygen saturations increased significantly (P<0.01) and the effective rate was 91.7%. CONCLUSION There are usually multiple obstruction levels in patients with severe OSAHS and the traditional surgical treatment is not ideal. This study demonstrated that with the assistance of coblation, combining CAUPPP with CCT for patients with severe OSAHS is an effective surgical method. It has less blood loss, minimally invasive, retained the normal function of the soft palate, etc., should be widely applied.
Adult ; Aged ; Catheter Ablation ; Female ; Humans ; Male ; Middle Aged ; Palate ; surgery ; Palate, Soft ; surgery ; Pharynx ; surgery ; Retrospective Studies ; Sleep Apnea, Obstructive ; surgery ; Tongue ; surgery ; Uvula ; surgery

Objective:Differences between randomized controlled trial (RCT) results and real world study (RWS) results may not represent a true efficacy-effectiveness gap because efficacy-effectiveness gap estimates may be biased when RWS and RCT differ significantly in study design or when there is bias in RWS result estimation. Secondly, when there is an efficacy- effectiveness gap, it should not treat every patient the same way but assess the real-world factors influencing the intervention's effectiveness and identify the subgroup likely to achieve the desired effect.Methods:Six databases (PubMed, Embase, Web of Science, CNKI, Wanfang Data, and VIP) were searched up to 31 st December 2022 with detailed search strategies. A scoping review method was used to integrate and qualitatively describe the included literature inductively. Results:Ten articles were included to discuss how to use the RCT research protocol as a template to develop the corresponding RWS research protocol. Moreover, based on correctly estimating the efficacy-effectiveness gap, evaluate the intervention effect in the patient subgroup to confirm the subgroup that can achieve the expected benefit-risk ratio to bridge the efficacy-effectiveness gap.Conclusion:Using real-world data to simulate key features of randomized controlled clinical trial study design can improve the authenticity and effectiveness of study results and bridge the efficacy-effectiveness gap.

Objective:Randomized controlled trials (RCT) usually have strict implementation criteria. The included subjects' characteristics of the conditions for the intervention implementation are quite different from the actual clinical environment, resulting in discrepancies between the risk-benefit of interventions in actual clinical use and the risk-benefit shown in RCT. Therefore, some methods are needed to enhance the extrapolation of RCT results to evaluate the real effects of drugs in real people and clinical practice settings.Methods:Six databases (PubMed, Embase, Web of Science, CNKI, Wanfang Data, and VIP) were searched up to 31 st December 2022 with detailed search strategies. A scoping review method was used to integrate and qualitatively describe the included literature inductively. Results:A total of 12 articles were included. Three methods in the included literature focused on: ①improving the design of traditional RCT to increase population representation; ②combining RCT Data with real-world data (RWD) for analysis;③calibrating RCT results according to real-world patient characteristics.Conclusions:Improving the design of RCT to enhance the population representation can improve the extrapolation of the results of RCT. Combining RCT data with RWD can give full play to the advantages of data from different sources; the results of the RCT were calibrated against real-world population characteristics so that the effects of interventions in real-world patient populations can be predicted.

Humans ; COVID-19/genetics* ; SARS-CoV-2/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; CRISPR-Cas Systems/genetics* ; RNA, Viral/genetics*