Primers and probes were designed based on the conserved regions of Senecavirus A(SVA),foot-and-mouth disease virus(FMDV),and porcine teschovirus(PTV)and used to devel-op a TaqMan fluorescent quantitative PCR method for detecting the above-mentioned three viru-ses.The triplex fluorescent quantitative PCR system was developed using recombinant positive plasmids containing conserved sequences of the three viruses as templates.After optimizing the conditions,the specificity,sensitivity,repeatability,standard curve,and mixed infection model were evaluated,and the constructed triplex fluorescent quantitative PCR was used for clinical detection.The results showed that this method could specifically detect SVA,FMDV and PTV without cross-reactivity with other pathogens with the minimal detection concentrations for SVA,FMDV,and PTV as low as 1X101 copies/μL,respectively.The coefficients of variation within and between groups were less than 5％.Furthermore,none of the three viruses were detected in 126 samples.The above results indicate that this method is highly specific,sensitive,and stable,making it suit-able for clinical detection.

Primers and probes were designed based on the conserved regions of Senecavirus A(SVA),foot-and-mouth disease virus(FMDV),and porcine teschovirus(PTV)and used to devel-op a TaqMan fluorescent quantitative PCR method for detecting the above-mentioned three viru-ses.The triplex fluorescent quantitative PCR system was developed using recombinant positive plasmids containing conserved sequences of the three viruses as templates.After optimizing the conditions,the specificity,sensitivity,repeatability,standard curve,and mixed infection model were evaluated,and the constructed triplex fluorescent quantitative PCR was used for clinical detection.The results showed that this method could specifically detect SVA,FMDV and PTV without cross-reactivity with other pathogens with the minimal detection concentrations for SVA,FMDV,and PTV as low as 1X101 copies/μL,respectively.The coefficients of variation within and between groups were less than 5％.Furthermore,none of the three viruses were detected in 126 samples.The above results indicate that this method is highly specific,sensitive,and stable,making it suit-able for clinical detection.

Background Sleep disturbances in children and adolescents have become a global public health concern,with pronounced issues in the Western China.Despite this,research on the prevalence of insomnia and its influencing factors among children and adolescents in Chongqing is still lacking.Objective To understand the prevalence of insomnia and analyze its influencing factors among children and adolescents in Chongqing,so as to provide references for clinical interventions and preventive strategies of insomnia.Methods A stratified random sampling method was used to select 9 969 children and adolescents in Chongqing from November to December 2021.Insomnia Severity Index(ISI)and a self-designed questionnaire on awareness of core mental health knowledge were administered.Binary Logistic regression analysis was used to identify the influencing factors of insomnia in this population.Results A total of 3 578 children and adolescents(35.89％)were found to have insomnia symptoms.There were statistically significant differences in the detection of insomnia symptoms among gender,ethnicity,education level,domicile,only-child status,parental education level,introversion/extroversion,parental relationship,average monthly per capita income,family history of mental illness,medical insurance coverage,living situation and understanding of mental health knowledge(P<0.05 or 0.01).Binary Logistic regression analysis identified several risk factors for insomnia,including female gender(OR=1.301,95％CI:1.192～1.419),ethnic minority status(OR=1.163,95％CI:1.015～1.333),junior high school for education level(OR=1.985,95％CI:1.774～2.220),senior high school for education level(OR=3.085,95％CI:2.749～3.462),non-only-child status(OR=1.127,95％CI:1.013～1.253),degree of harmony between parents is not high or not harmonious[(OR=1.846,95％CI:1.669～2.041 for relatively harmonious;OR=2.524,95％CI:2.214～2.877 for generally harmonious;OR=2.452,95％CI:1.999～3.007 for not very harmonious;OR=2.926,95％CI:2.307～3.710 for very discordant)]and incomplete medical insurance coverage(OR=1.218,95％CI:1.093～1.358).Protective factors included an extroverted personality(OR=0.838,95％CI:0.766～0.917),absence of family history of mental illness(OR=0.719,95％CI:0.549～0.941),junior high school for mother's education level(OR=0.822,95％CI:0.734～0.920),senior high school or secondary specialized school for mother's education level(OR=0.862,95％CI:0.752～0.988),college and above for mother's education level(OR=0.748,95％CI:0.633～0.884)and knowledge of mental health(OR=0.854,95％CI:0.778～0.937).Conclusion The problem of insomnia among children and adolescents in Chongqing is quite serious.Risk factors for insomnia include female gender,ethnic minority status,higher education levels,being a non-only child,poor family relationships and incomplete medical insurance.Conversely,an extroverted personality,absence of family history of mental illness,higher maternal education and knowledge of mental health serve as protective factors against insomnia.

As precious Chinese medicinal materials， Moschus and Bovis Calculus are often used in the treatment of acute and severe patients. However， due to their scarce sources and high prices， wild animal resources are in urgent need of protection， natural Moschus and Bovis Calculus can no longer meet the needs of preparation production. The use of substitutes such as Moschus Artifactus， Bovis Calculus Artifactus and Bovis Calculus Sativus has alleviated the shortage of original medicinal materials to some extent， and has been widely used in the production and use of traditional Chinese medicine （TCM） preparations. According to statistics， the 2020 edition of Chinese Pharmacopoeia contains 75 TCM preparations containing Moschus and 95 preparations containing Bovis Calculus， but in the quality standards of these 134 TCM preparations， the difference between natural medicinal materials and their substitutes is not very obvious， and some quality control projects are relatively simple. Based on this， the author intends to sort out the quality standards of TCM preparations containing Moschus or Bovis Calculus in the 2020 edition of Chinese Pharmacopoeia （volume Ⅰ）， including the type， drug form， prescription dosage， maximum daily （time） dosage and the quality control items of Moschus and Bovis Calculus in the preparation， in order to explore the rationality of the quality standard of TCM preparations containing Moschus or Bovis Calculus， and to give some suggestions on standardizing the use types of Moschus and Bovis Calculus， improving the quality control items of Moschus and Bovis Calculus in TCM preparations combining modern research achievements and advanced technology.

Genome-wide physical protein-protein interaction(PPI)mapping remains a major chal-lenge for current technologies.Here,we reported a high-efficiency BiFC-seq method,yeast-enhanced green fluorescent protein-based bimolecular fluorescence complementation(yEGFP-BiFC)coupled with next-generation DNA sequencing,for interactome mapping.We first applied yEGFP-BiFC method to systematically investigate an intraviral network of the Ebola virus.Two-thirds(9/14)of known interactions of EBOV were recaptured,and five novel interactions were discovered.Next,we used the BiFC-seq method to map the interactome of the tumor protein p53.We identified 97 interactors of p53,more than three-quarters of which were novel.Furthermore,in a more complex background,we screened potential interactors by pooling two BiFC libraries together and revealed a network of 229 interactions among 205 proteins.These results show that BiFC-seq is a highly sensitive,rapid,and economical method for genome-wide interactome map-ping.

Objective:To research the quality of postoperative recovery of patients with lung cancer through the(PQRS),and compare the recovery quality after video-assisted thoracoscopic surgery (VATS) or traditional open lobectomy.Method:PQRS scale was used to evaluate the recovery of 140 patients with lung lesions preoperatively and postoperative 1 day,3 days,7 days,14 days and 1 month.95 patients who met the set standard and complete the PQRS scales were enrolled and divided into video-assisted thoracoscope group or traditional thoracotomy group.This article mainly compared and analysied the quality of postoperative recovery of patients in both groups.Result:Except the anesthesia time,other general datum showed no statistical difference.In total recovery rate,the video-assisted thoracoscope group has significant difference when compared with the traditional thoracotomy group(P ＜ 0.05).The recovery rates of VATS group in feeling of the harmful factors,emotional factors and the daily life are higher than that of the traditional thoracotomy group,with statistically significant differences(P ＜0.05).However,the recovery rates in the physiological factors and cognitive factors had no statistical difference between two groups(P＞ 0.05).Conclusion:PQRS can effectively evaluate the quality of postoperative recovery in patients with lung cancer,and VATS is helpful to quick recovery postoperatively.

Objective To investigate the effect of electroacupuncture at points Xiaohai and Xiajuxu on the expressions of tumour necrosis factor-α (TNF-α) and choline acetyltransferase (ChAT) in serum and nicotinic acetylcholine receptorα 7 (α 7 nAchR) in duodenal tissues in a rat model of duodenal ulcer (DU) and preliminarily explore the relative specificity of He-Sea point in “treating visceral diseases with He-Sea point”.Methods Forty healthy SD rats were randomized into blank (A), model (B), Xiaohai (C) and Xiajuxu (D) groups, 10 rats each. A rat model of DU was made by subcutaneous injection of 10% cysteamine hydrochloride at the right buttock. After successful model making, group C was given electroacupuncture at point Xiaohai and group D, at point Xiajuxu. Duodenal tissue ulcer was macroscopically observed and scored in every group of rats. Rat serum expression of TNF-α was determined by double antibody sandwich enzyme-linked immunosorbent assay (ELISA); rat serum expression of ChAT, by ultraviolet spectrophotometry & colorimetry; rat duodenal expression ofα7 nAchR, by Western blot.Results After model making, the duodenal ulcer score was significantly higher in groups B, C and D than in group A (allP<0.01) and significantly lower in groups C and D than in group B (bothP<0.01) and in group D than in group C (P<0.01). TNF-α expression was significantly higher in group B than in group A (P<0.01) and significantly lower in groups C and D than in group B (bothP<0.01) and in group D than in group C (P<0.01). ChAT expression tended to increase in group C compared with group B but there was no statistically significant difference (P>0.05) and was significantly higher in group D than in group B (P<0.01) or C (P<0.05).α7 nAchR expression was significantly higher in groups C and D than in group B (bothP<0.01). There was a positive correlation between ChAT andα7 nAchR expressions in every group (r=0.444,P=0.007).Conclusions Electroacupuncture at both points Xiaohai and Xiajuxu can reduce the duodenal ulcer score and serum TNF-α expression and increase serum ChAT and duodenalα7 nAchR expressions in DU rats. The results show that the therapeutic effect of electroacupuncture on duodenal ulcer may be produced by regulating TNF-α. Its mechanism may be activating cholinergic anti-inflammatory pathway to produce an anti-inflammatory effect. The effect being better in group D than in group C suggests that Xiajuxu has the relative specificity.