In order to determine the causative agents responsible for the lethality of tumor disease in breeders,liver,spleen and other tissues of dead chickens were collected,and pathogen detection,se-quencing and genetic evolution analysis were carried out by PCR.The results showed that all three samples were negative for reticuloendotheliosis virus(REV),samples 1 and 2 wereavian leukosis virus subgroup J(ALV-J)positive,and sample 3 was positive for ALV-J and Marek's disease virus(MDV),which preliminarily determined that the breeder died of ALV-J infection or ALV-J and MDV mixed infection.Among them,the ALV-J gp85 gene had the highest nucleotide homology with the reference strain of ALV-J,ranging from 87.70％to 99.30％,and was in the same branch with the reference strain of subgroup J.Amino acid homology ranged from 78.00％to 96.20％,with some mutations.The nucleotide homology between the MDV meq gene and the vv+strain was the highest,which was 98.00％-99.00％,and they were in the same branch.The homology of amino acids was 95.90％-98.20％,and there were multiple mutation sites,among which the 176th amino acid destroyed the original Pro repeat of the virus MDV meq due to the mutation of Pro to Arg or Ala,which may lead to the enhancement of its virulence.This study will provide a reference for the prevention and control of neoplastic diseases in breeders in this region.

Objective To investigate the effect of Lianpu Yin on duodenal microinflammation in rats with functional dyspepsia(FD)by regulating NLRP3 activation.Methods Wistar rats were randomly divided into blank group and model group.FD rats were reconstructed by iodoacetamide method(2%sucrose solution containing 0.1%iodoacetamide),and the model was verified.FD model rats were randomly divided into model group,Lianpu Yin group and Moxapride group by random number expression method.After a period of two weeks of administration,measurements were taken to determine the body mass,three-hour food consumption,as well as the rates of gastric emptying and intestinal propulsion.The pathological structure of duodenal tissue was observed by HE staining.The serum levels of IL-1β and IL-18 were quantified using the enzyme-linked immunosorbent assay(ELISA)method.The expression levels of NLRP3 and Caspase-1 in each group were detected by Western blot.Expression levels of NLRP3 and Caspase-1 proteins were detected by immunofluorescence.Results Compared with the blank group,body weight,food intake at 3 h,gastric emptyand intestinal propulsion rate in model group were significantly decreased(P<0.01),and inflammatory infiltration of duodenum tissue appeared in the model group.Meanwhile,the expressions of NLRP3 and Caspase-1 proteins,as well as the levels of IL-1β and IL-18 in the duodenal tissue of the model group,showed significant increasing(P<0.05).Compared with the model group,rats in the Lianpu Yin and Moxapride groups displayed significant increasing in body weight,gastric emptying rate,and intestinal propulsion rate(P<0.01).Additionally,inflammatory infiltration of duodenum tissue reduced in these groups.Furthermore,NLRP3 and Caspase-1 protein expressions,as well as IL-1β and IL-18 levels,significantly decreased in the Lianpu Yin and Moxapride groups compared to the model group(P<0.05).Conclusion Lianpu Yin can treat FD rats by inhibiting duodenal microinflammation and then restoring gastrointestinal motility,which may be related to the abnormal activation of NLRP3 inflammasome.

In order to determine the causative agents responsible for the lethality of tumor disease in breeders,liver,spleen and other tissues of dead chickens were collected,and pathogen detection,se-quencing and genetic evolution analysis were carried out by PCR.The results showed that all three samples were negative for reticuloendotheliosis virus(REV),samples 1 and 2 wereavian leukosis virus subgroup J(ALV-J)positive,and sample 3 was positive for ALV-J and Marek's disease virus(MDV),which preliminarily determined that the breeder died of ALV-J infection or ALV-J and MDV mixed infection.Among them,the ALV-J gp85 gene had the highest nucleotide homology with the reference strain of ALV-J,ranging from 87.70％to 99.30％,and was in the same branch with the reference strain of subgroup J.Amino acid homology ranged from 78.00％to 96.20％,with some mutations.The nucleotide homology between the MDV meq gene and the vv+strain was the highest,which was 98.00％-99.00％,and they were in the same branch.The homology of amino acids was 95.90％-98.20％,and there were multiple mutation sites,among which the 176th amino acid destroyed the original Pro repeat of the virus MDV meq due to the mutation of Pro to Arg or Ala,which may lead to the enhancement of its virulence.This study will provide a reference for the prevention and control of neoplastic diseases in breeders in this region.

Objective To investigate the effect of Lianpu Yin on duodenal microinflammation in rats with functional dyspepsia(FD)by regulating NLRP3 activation.Methods Wistar rats were randomly divided into blank group and model group.FD rats were reconstructed by iodoacetamide method(2%sucrose solution containing 0.1%iodoacetamide),and the model was verified.FD model rats were randomly divided into model group,Lianpu Yin group and Moxapride group by random number expression method.After a period of two weeks of administration,measurements were taken to determine the body mass,three-hour food consumption,as well as the rates of gastric emptying and intestinal propulsion.The pathological structure of duodenal tissue was observed by HE staining.The serum levels of IL-1β and IL-18 were quantified using the enzyme-linked immunosorbent assay(ELISA)method.The expression levels of NLRP3 and Caspase-1 in each group were detected by Western blot.Expression levels of NLRP3 and Caspase-1 proteins were detected by immunofluorescence.Results Compared with the blank group,body weight,food intake at 3 h,gastric emptyand intestinal propulsion rate in model group were significantly decreased(P<0.01),and inflammatory infiltration of duodenum tissue appeared in the model group.Meanwhile,the expressions of NLRP3 and Caspase-1 proteins,as well as the levels of IL-1β and IL-18 in the duodenal tissue of the model group,showed significant increasing(P<0.05).Compared with the model group,rats in the Lianpu Yin and Moxapride groups displayed significant increasing in body weight,gastric emptying rate,and intestinal propulsion rate(P<0.01).Additionally,inflammatory infiltration of duodenum tissue reduced in these groups.Furthermore,NLRP3 and Caspase-1 protein expressions,as well as IL-1β and IL-18 levels,significantly decreased in the Lianpu Yin and Moxapride groups compared to the model group(P<0.05).Conclusion Lianpu Yin can treat FD rats by inhibiting duodenal microinflammation and then restoring gastrointestinal motility,which may be related to the abnormal activation of NLRP3 inflammasome.

ObjectiveTo examine the inhibitory effects of berberine compounds， including columbamine， on acetylcholinesterase from the perspectives of drug-target binding affinity and kinetics and explore the blood-brain barrier （BBB） permeability of these compounds in different multi-component backgrounds. MethodThe median inhibitory concentration （IC₅₀） of acetylcholinesterase by berberine compounds including columbamine was measured using the Ellman-modified spectrophotometric method. The binding kinetic parameters （K_off） of these compounds with acetylcholinesterase were determined using the enzyme activity recovery method. A qualitative analysis of the ability of these components to penetrate the BBB and arrive at the brain tissue in diverse multi-component backgrounds （including medicinal herbs and compound formulas） was conducted using ultra performance liquid chromatography-high resolution mass spectrometry （UPLC-HRMS）. ResultBerberine compounds， including columbamine， exhibited strong inhibition of acetylcholinesterase， with IC₅₀ values in the nanomolar range. Moreover， they displayed better drug-target binding kinetics characteristics （with smaller K_off values） than the positive control of donepezil hydrochloride （P<0.01）， indicating a longer inhibition duration of acetylcholinesterase. Berberine components such as columbamine could penetrate the BBB to arrive at brain tissue in the form of a monomer， as well as in the multi-component backgrounds of Coptis and Phellodendri Chinensis Cortex medicinal extracts and the compound formula Huanglian Jiedutang. ConclusionThese berberine compounds such as columbamine exhibit a strong inhibitory effect on acetylcholinesterase and can arrive at brain tissue in multi-component backgrounds. In the level of pharmacological substance， this supports the clinical efficacy of compound Huanglian Jiedutang in improving Alzheimer's disease， providing data support for elucidating the pharmacological basis of compound Huanglian Jiedutang.

Objective:To retrospectively analyze the results of second-trimester serological prenatal screening and explore the factors which may influence the false-positive rate (FPR).Methods:From January 2013 to December 2022, false-positive samples with follow-up outcomes from 632, 825 second-trimester serological prenatal screening samples tested at Sichuan Provincial Maternity and Child Health Care Hospital were selected as the study group, while true-negative samples were 1 : 1 matched as the control group by propensity-score matching (PSM). Univariate and Multivariate Logistic Regression Models were used to analyze the influencing factors. The study has approved by the Medical Ethics Committee of the Sichuan Provincial Maternity and Child Health Care Hospital (Ethic No.20240607-270).Results:The study and control groups were each matched with 305, 998 cases. Univariate analysis showed that sampling season, the difference between ultrasound and gestational weeks calculated by last menstrual period (LMP), monthly median multiple of the median (mMoM) of alpha-fetoprotein (AFP), and monthly mMoM of free β-human chorionic gonadotropin (free β-hCG) were significantly different between the two groups ( P<0.05). Multivariate Logistic Regression analysis showed that winter ( OR=0.938; 95% CI: 0.893~0.985), monthly AFP mMoM ≥ 1.11 ( OR=0.846; 95% CI: 0.761~0.941), monthly free β-hCG mMoM ≤ 0.89 ( OR=0.827; 95% CI: 0.737~0.929) are protective factors for FPR increase, whilst spring ( OR=1.124; 95% CI: 1.072~1.179), summer ( OR=1.121; 95% CI: 1.062~1.183), the difference between ultrasound and gestational weeks calculated by LMP of 8~14 days ( OR=1.319; 95% CI: 1.241~1.402), > 14 days ( OR=1.689; 95% CI: 1.542~1.850), monthly AFP mMoM of 0.90~0.94 ( OR=1.088; 95% CI: 1.046~1.131), and monthly free β-hCG mMoM of 1.05~1.10 ( OR=1.046; 95% CI: 1.000~1.094), ≥ 1.11 ( OR=1.062; 95% CI: 1.002~1.126) are risk factors for FPR increase. Conclusion:Sampling season, difference between ultrasound and gestational weeks by LMP, monthly AFP mMoM, and monthly free β-hCG mMoM are risk factors for FPR during serological prenatal screening. Screening laboratories should look for the cause of abnormal FPR through such factors and adjust them accordingly.

Objective Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)and network pharmacology technology combined with pharmacodynamic experiments were used to analyze the quality markers(Q-markers)in Saorilao Qingfei Zhike Capsules.Methods Using UPLC-Q-TOF-MS technology,the chemical components in different polar extracts of Saorilao Qingfei Zhike Capsules was analyzed.Potential pharmacological components were screened by using antitussive and expectorant models.The"components-targets-diseases"network was constructed and potential Q-markers were screened by network pharmacology technology.Then we conducted pharmacodynamic validation to confirm the Q-markers,which have antitussive and expectorant effects in Saorilao Qingfei Zhike Capsules.Results A total of 120 compounds were obtained from the Saorilao Qingfei Zhike Capsules through qualitative analysis.Among the extracts of different polarity,44 compounds were derived from petroleum ether extract,85 compounds were derived from ethyl acetate extract,79 compounds were derived from n-butanol extract,and 71 compounds were derived from water extract.The results of pharmacological experiments showed that among extracts of different polarity,petroleum ether extract had the best antitussive effect,while n-butanol extract had the best expectorant effect.Three core components for eliminating phlegm and relieving cough were screened through network pharmacology techniques:farcalinol,farcalinediol,and rubimaillin.Pharmacodynamic studies verified that all core components mentioned above have certain antitussive and expectorant effects.Conclusion Based on the above research,farcalinol,farcalindiol,and rubimaillin can be used as Q-markers for the antitussive and expectorant effects of Saorilao Qingfei Zhike Capsules.This paper provides reference for the quality standard of Saorilao Qingfei Zhike Capsules.

The occurrence of benign prostate hyperplasia(BPH)was related to disrupted sex steroid hormones,and metformin(Met)had a clinical response to sex steroid hormone-related gynaecological disease.How-ever,whether Met exerts an antiproliferative effect on BPH via sex steroid hormones remains unclear.Here,our clinical study showed that along with prostatic epithelial cell(PEC)proliferation,sex steroid hormones were dysregulated in the serum and prostate of BPH patients.As the major contributor to dysregulated sex steroid hormones,elevated dihydrotestosterone(DHT)had a significant positive rela-tionship with the clinical characteristics of BPH patients.Activation of adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)by Met restored dysregulated sex steroid hormone homeostasis and exerted antiproliferative effects against DHT-induced proliferation by inhibiting the formation of androgen receptor(AR)-mediated Yes-associated protein(YAP1)-TEA domain transcription factor(TEAD4)heterodimers.Met's anti-proliferative effects were blocked by AMPK inhibitor or YAP1 over-expression in DHT-cultured BPH-1 cells.Our findings indicated that Met would be a promising clinical therapeutic approach for BPH by inhibiting dysregulated steroid hormone-induced PEC proliferation.

Objective To investigate the effect of the sling massage exercise on postpartum pelvic girdle pain(PPGP). Methods From March,2022 to May,2023,56 patients with PPGP in Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine were randomly divided into control group(n=28)and observation group(n=28).Both groups received routine physical therapy and healthy education,while the control group received exer-cise therapy and the observation group received sling massage exercise,for six weeks.They were assessed with Visual Analog Scale(VAS),Pelvic Girdle Questionnaire(PGQ)and active straight leg raise(ASLR)test before and after treatment,and measured resting and contraction thickness of transversus abdominis with ultrasound;and integrated electromyography(iEMG),root mean square(RMS),median frequency(MF)and mean power fre-quency(MPF)of pelvic floor muscles with surface electromyography. Results After treatment,the total effective rate was 96.43%in the observation group and 78.57%in the control group(Z=2.728,P<0.05).All the indexes improved in both groups(|t|>2.080,P<0.05)except resting thickness of transverse abdominis muscle,and improved more in the observation group than in the control group(|t|>2.161,P<0.05). Conclusion The sling massage exercise can alleviate pain,enhance the stability of the core muscle group,and improve load transfer and quality of life,better than exercise therapy.

Objective To explore the effect of sling exercise with Tuina therapy on kinesiophobia in old patients with lumbar disc herniation,and analyze the mechanism based on brain-bone axis. Methods A total of 56 old patients with chronic lumbar disc herniation and kinesiophobia were selected from the Reha-bilitation Hospital of the Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine from September,2022 to December,2023;and randomly divided into control group(n=28)and experimental group(n=28).The control group accepted conventional exercise therapy,while the experimental group accepted sling exercise with Tuina therapy,for four weeks.They were assessed with simplified Chinese version of Tampa Scale of Kinesiophobia(TSK),Japanese Orthopaedic Association score(JOA)and Visual Analogue Scale for pain(VAS)before and after treatment,while the bone mineral density(BMD)was tested,the levels of osteoprote-gerin(OPG),norepinephrine(NE)and corticosteroids(Cor)in serum were measured,and the median frequency(MF)of weak-link erector spinae was detected with surface electromyography. Results Two cases dropped off in the control group,and one in the experimental group.The scores of all the assessment improved in both groups after treatment(|t|>14.168,P<0.001),as well as the serum levels of OPG,NE and Cor(|t|>2.103,P<0.05),BMD(|t|>2.726,P<0.05),and MF of erector spinae(|t|>14.736,P<0.001);all of them were better in the experimental group than in the control group(|t|>2.154,P<0.05). Conclusion Sling exercise with Tuina therapy can improve the pain and kinesiophobia of lumbar disc herniation in the old adults,which may promote the recovery of physical and mental function through regulating the levels of hor-mones and neurotransmitters related to the brain-bone axis.