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.

Objective:To compare the application of cloning sequencing and third-generation nanopore sequencing technology in the identification of raw materials mixed in Tibetan patent medicine Shiliujianwei powder,and to pro-vide a reference for the establishment of the specific identification method for the formulation of medicinal prepara-tions using the raw powder of medicinal herbs or herbal pieces.Methods:By investigating the different concentra-tions of ExTaq enzyme,genomic DNA,upstream and downstream primers in the PCR amplification system,the suit-able PCR amplification system of genomic DNA universal primers for self-made Tibetan medicine Shiliujianwei powder was determined.The amplified products of Shiliujianwei powder were sequenced by two methods,the first was cloned and sequenced by Sanger sequencing technology,and the second was sequenced by third generation nanopore sequen-cing technology.Results:The addition of ExTaq enzyme,upstream and downstream primers and genomic DNA in 20 μL PCR amplification system of Shiliujianwei powder and single medicinal samples were determined to be 0.4,1.5 and 1 μL,respectively.The amplification products of the DNA from self-made Shiliujianwei powder were cloned and sequenced by Sanger sequencing technology,then only the ITS2 sequence of Carthami flos was obtained.The amplified products of self-made pomegranate Jianwei powder DNA were sequenced by three-generation nanopore se-quencing technology,and the ITS2 sequences of Granati semen,Carthami flos,Piperis longi fructus and Amomi fruc-tus rotundus were finally obtained,but the ITS2 sequence of cinnamomi cortex was not obtained.Conclusion:Both cloning sequencing and the third generation nanopore sequencing could solve the problem of overlapping peaks in the direct sanger sequencing for the universal primer amplification products of patent drugs.The third generation nano-pore sequencing was better than cloning sequencing in the sequencing of the universal primer amplification products of patent drugs,but there were still one raw material medicine that have not been detected.It is of certain reference val-ue for the molecular biological identification and DNA barcoding study of different raw materials in the patent medi-cine to establish the specific identification method of Tibetan patent medicine Shiliujianwei powder.

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 explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg^-1·d^-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg^-1·d^-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg^-1·d^-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg^-1·d^-1） + amoxicillin （303 mg·kg^-1·d^-1） + clarithromycin （151.67 mg·kg^-1·d^-1） + colloidal pectin capsules （30.3 mg·kg^-1·d^-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.

ObjectiveTo explore the effect of modified Lianpoyin formula （LPYJWF） in the treatment of Helicobacter pylori （Hp）-associated gastric mucosal damage based on mitochondrial autophagy and NLRP3 inflammasome signaling pathway. MethodsA total of 60 eight-week-old Balb/c male mice were assigned via the random number table method into control， model， high-dose LPYJWF （LPYJWF-H， 27.3 g·kg^-1·d^-1）， medium-dose LPYJWF （LPYJWF-M， 13.65 g·kg^-1·d^-1）， low-dose LPYJWF （LPYJWF-L， 6.83 g·kg^-1·d^-1）， and quadruple therapy groups. Except the control group， other groups were modeled for Hp infection. Mice were administrated with LPYJWF at corresponding doses by gavage. Quadruple therapy group was given omeprazole （6.06 mg·kg^-1·d^-1） + amoxicillin （303 mg·kg^-1·d^-1） + clarithromycin （151.67 mg·kg^-1·d^-1） + colloidal pectin capsules （30.3 mg·kg^-1·d^-1） by gavage. The control group was given an equal volume of 0.9% NaCl for 14 days. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of gastric mucosa， and Warthin-Starry （W-S） silver staining was used to detect Hp colonization. Transmission electron microscopy was employed to observe the mitochondrial ultrastructure of the gastric tissue， and immunofluorescence co-localization assay was adopted to detect the expression of mitochondrial transcription factor A （TFAM） and translocase of the outer mitochondrial membrane member 20 （TOMM20）. The water-soluble tetrazolium salt method and thiobarbituric acid method were used to determine the levels of superoxide dismutase （SOD） and malondialdehyde （MDA）， respectively， in the gastric tissue. Western blot was employed to measure the protein levels of PTEN-induced kinase 1 （PINK1）， Parkin， p62， microtubule-associated protein 1 light chain 3 （LC3）， NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18）. Real-time quantitative PCR was employed to assess the mRNA levels of PINK1， Parkin， p62， and LC3. ResultsCompared with the control group， the model group presented obvious gastric mucosal damage， colonization of a large number of Hp， severe mitochondrial damage， vacuolated structures due to excessive autophagy， reduced TOMM20 and TFAM co-expression in the gastric mucosal tissue， and reduced SOD and increased MDA （P<0.01）. In addition， the gastric tissue in the model group showed up-regulated protein and mRNA levels of PINK1， Parkin， and LC3 and down-regulated protein and mRNA levels of p62 （P<0.01， as well as increased expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 （P<0.01）. Compared with the model group， the LPYJWF and quadruple therapy groups showed alleviated pathological damage of gastric mucosa， reduced Hp colonization， mitigated mitochondrial damage， and increased co-expression of TOMM20 and TFAM. The SOD level was elevated in the LPYJWF-L group （P<0.01）， and the MDA levels became lowered in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. Furthermore， the LPYJWF and quadruple therapy groups showed down-regulated mRNA levels of PINK1， Parkin， and LC3 and protein levels of PINK1 and Parkin， and up-regulated mRNA level of p62 （P<0.01）. The LPYJWF-M， LPYJWF-H， and quadruple therapy groups showcased down-regulated LC3 Ⅱ/LC3 Ⅰ level （P<0.05， P<0.01） and up-regulated protein level of p62 （P<0.01）. The expression of inflammasome-associated proteins NLRP3， ASC， IL-1β， and IL-18 were reduced in the LPYJWF and quadruple therapy groups （P<0.05， P<0.01）. ConclusionLPYJWF ameliorates gastric mucosal damage and exerts mucosa-protective effects in Hp-infected mice， which may be related to the inhibition of excessive mitochondrial autophagy， thereby inhibiting the activation of the NLRP3 inflammasome pathway.

Objective:To explore a systematic training for quick identification for diagnosis and treatment of the causes of reversible cardiac arrest,which was assisted by bedside ultrasound simulator combined with memory mnemonic,so as to improve the treatment skill of physicians for the causes of disease,and enhance the success rate of cardiopulmonary resuscitation.Methods:Thirty clinical resident doctors who were rotating in the emergency intensive care unit(ICU)of Xuanwu Hospital,Capital Medical University,from May 2023 to May 2024 were selected.The bedside ultrasound simulator combined with memory mnemonic was used to simulate training,which can improve the ability of resident doctors in quickly determining the cause of reversible cardiac arrest.The self-made questionnaire was used to investigate the baseline situation of resident doctors before they were trained,and the teaching effectiveness after they received training.After training,the grasping states of resident doctors for the cause of reversible cardiac arrest were detected through 10 kinds of clinical cases included pulmonary embolism,myocardial infarction,aortic dissection,pericardial tamponade,ventricular fibrillation,electrolyte disturbance,hypovolemic shock,tension pneumothorax,poisoning and hypothermia.Results:The duration of quick retelling of resident doctors for the causes of reversible cardiac arrest was(15.07±3.68)s after training,which was lower than(47.27±10.25)s before training,and the difference was significant(t=14.763,P<0.05).The awareness rate of them for the causes of reversible cardiac arrest was 100%after training,which was significantly higher than 3.33%before training,and the difference was significant(x2=56.129,P<0.05).The identification of resident doctor for ultrasonic image was zero basis before training,and the identification rate was 0%.All of resident doctors can achieve 100%identification for routine ultrasound,cardiac arrest,pericardial tamponade and ventricular fibrillation within the simulator after training,and the correct recognition rates of them for pulmonary embolism,tension pneumothorax,acute myocardial infarction and aortic dissection were respectively 86.67%,63.33%,70%and 66.67%.After training,the duration of correct recognition of resident doctors for pulmonary embolism,tension pneumothorax and acute myocardial infarction were respectively(58.77±19.61)s,(69.05±10.47)s and(75.52±10.51)s,all of which were less than the duration of incorrect recognition,and the differences were statistically significant(t=2.153,2.781,2.124,P<0.05).Conclusion:Ultrasound combined with memory mnemonic can help resident doctors to quickly establish the clinical thinking clue about hypoxia,hypovolemia,hypo/hyperkalemia,hypo/hyperthermia,hypo/hyperglycemia,tamponade cardiac,thrombosis pulmonary,thrombosis coronary,toxins,tension pneumothorax(5H5T)for the cause of reversible cardiac arrest,which will contribute to strengthen and improve the practical ability of clinician in quick judgement for reversible etiology.

Objective:To compare the application of cloning sequencing and third-generation nanopore sequencing technology in the identification of raw materials mixed in Tibetan patent medicine Shiliujianwei powder,and to pro-vide a reference for the establishment of the specific identification method for the formulation of medicinal prepara-tions using the raw powder of medicinal herbs or herbal pieces.Methods:By investigating the different concentra-tions of ExTaq enzyme,genomic DNA,upstream and downstream primers in the PCR amplification system,the suit-able PCR amplification system of genomic DNA universal primers for self-made Tibetan medicine Shiliujianwei powder was determined.The amplified products of Shiliujianwei powder were sequenced by two methods,the first was cloned and sequenced by Sanger sequencing technology,and the second was sequenced by third generation nanopore sequen-cing technology.Results:The addition of ExTaq enzyme,upstream and downstream primers and genomic DNA in 20 μL PCR amplification system of Shiliujianwei powder and single medicinal samples were determined to be 0.4,1.5 and 1 μL,respectively.The amplification products of the DNA from self-made Shiliujianwei powder were cloned and sequenced by Sanger sequencing technology,then only the ITS2 sequence of Carthami flos was obtained.The amplified products of self-made pomegranate Jianwei powder DNA were sequenced by three-generation nanopore se-quencing technology,and the ITS2 sequences of Granati semen,Carthami flos,Piperis longi fructus and Amomi fruc-tus rotundus were finally obtained,but the ITS2 sequence of cinnamomi cortex was not obtained.Conclusion:Both cloning sequencing and the third generation nanopore sequencing could solve the problem of overlapping peaks in the direct sanger sequencing for the universal primer amplification products of patent drugs.The third generation nano-pore sequencing was better than cloning sequencing in the sequencing of the universal primer amplification products of patent drugs,but there were still one raw material medicine that have not been detected.It is of certain reference val-ue for the molecular biological identification and DNA barcoding study of different raw materials in the patent medi-cine to establish the specific identification method of Tibetan patent medicine Shiliujianwei powder.

Objective:To explore a systematic training for quick identification for diagnosis and treatment of the causes of reversible cardiac arrest,which was assisted by bedside ultrasound simulator combined with memory mnemonic,so as to improve the treatment skill of physicians for the causes of disease,and enhance the success rate of cardiopulmonary resuscitation.Methods:Thirty clinical resident doctors who were rotating in the emergency intensive care unit(ICU)of Xuanwu Hospital,Capital Medical University,from May 2023 to May 2024 were selected.The bedside ultrasound simulator combined with memory mnemonic was used to simulate training,which can improve the ability of resident doctors in quickly determining the cause of reversible cardiac arrest.The self-made questionnaire was used to investigate the baseline situation of resident doctors before they were trained,and the teaching effectiveness after they received training.After training,the grasping states of resident doctors for the cause of reversible cardiac arrest were detected through 10 kinds of clinical cases included pulmonary embolism,myocardial infarction,aortic dissection,pericardial tamponade,ventricular fibrillation,electrolyte disturbance,hypovolemic shock,tension pneumothorax,poisoning and hypothermia.Results:The duration of quick retelling of resident doctors for the causes of reversible cardiac arrest was(15.07±3.68)s after training,which was lower than(47.27±10.25)s before training,and the difference was significant(t=14.763,P<0.05).The awareness rate of them for the causes of reversible cardiac arrest was 100%after training,which was significantly higher than 3.33%before training,and the difference was significant(x2=56.129,P<0.05).The identification of resident doctor for ultrasonic image was zero basis before training,and the identification rate was 0%.All of resident doctors can achieve 100%identification for routine ultrasound,cardiac arrest,pericardial tamponade and ventricular fibrillation within the simulator after training,and the correct recognition rates of them for pulmonary embolism,tension pneumothorax,acute myocardial infarction and aortic dissection were respectively 86.67%,63.33%,70%and 66.67%.After training,the duration of correct recognition of resident doctors for pulmonary embolism,tension pneumothorax and acute myocardial infarction were respectively(58.77±19.61)s,(69.05±10.47)s and(75.52±10.51)s,all of which were less than the duration of incorrect recognition,and the differences were statistically significant(t=2.153,2.781,2.124,P<0.05).Conclusion:Ultrasound combined with memory mnemonic can help resident doctors to quickly establish the clinical thinking clue about hypoxia,hypovolemia,hypo/hyperkalemia,hypo/hyperthermia,hypo/hyperglycemia,tamponade cardiac,thrombosis pulmonary,thrombosis coronary,toxins,tension pneumothorax(5H5T)for the cause of reversible cardiac arrest,which will contribute to strengthen and improve the practical ability of clinician in quick judgement for reversible etiology.

Objective:To explore the clinical application value of DNA image cytometry ploidy analysis (DNA-ICM) in the pathological diagnosis of malignant pleural effusion.Methods:A retrospective case series study was conducted. The clinical data of 101 patients with pleural effusion from October to December 2021 in Shanxi Bethune Hospital were retrospectively analyzed. Liquid-based cytology (LBC) and DNA-ICM were performed on pleural effusion specimens. The sensitivity and specificity of the two methods were compared with the clinical diagnosis, imaging, biopsy, and follow-up results of the patients.Results:Among the pleural effusions of 101 patients, 39 were malignant pleural effusions and 62 were benign pleural effusions. The sensitivity of LBC and DNA-ICM in diagnosing malignant tumor cells in pleural effusions was 74.7% and 94.9%, respectively, and the specificity was 98.4% and 83.9%, respectively; the combination of the two had an increased diagnostic positivity rate compared with that of LBC alone [36.6% (37/101) vs. 28.7% (29/101)]. Seven cases with positive DNA-ICM but negative LBC result were followed up, and 1 case was diagnosed as small cell lung cancer. Conclusions:DNA-ICM can effectively improve the positive cytology detection rate of pleural effusion, and the combined detection of DNA-ICM and LBC can reduce the underdiagnosis rate of cytology, which is of great clinical value in the pathological diagnosis of malignant pleural effusion.

Objective To investigate the clinical efficacy and postoperative aortic changes of 3D printing-assisted stent with in-vitro pre-fenestration technique in the treatment of complex type B aortic dissection.Methods A retrospective analysis was conducted on 28 patients with complex type B aortic dissection who underwent thoracic endovascular aortic repair(TEVAR)using stent with in-vitro pre-fenestration technique.The patients were divided into two groups based on the methods of stent with in-vitro pre-fenestration:the 3D printing-assisted stent pre-fenestration group(3D printing group)consisting of 13 patients and traditional stent pre-fenestration group(traditional group)consisting of 15 patients.Various parameters,including hospital duration,surgical duration,duration of stent pre-fenestration in vitro,dose of contrast agent,intraoperative blood loss,immediate postoperative internal leakage rate,30 d postoperative internal leakage rate,30 d postoperative mortality,and the change rate of true and false lumen diameter[the change rate of true and false lumen diameter=(postoperative true and false lumen diameter-preoperative true and false lumen diameter)/preoperative true and false lumen diameter× 100％],were compared between the two groups.Results Compared with the traditional group,the 3D printing group exhibited a considerable reduction in surgical duration[(148.46±27.20)min vs(175.46±22.04)min,P＜0.05].Additionally,the 30 d postoperative type Ⅰ internal leakage rate was lower in the 3D printing group(X2=4.044,P=0.044).Moreover,the change(dilation)rate of true lumen diameter in stent coverage section showed a notable increase in the 3D printing group(P＜0.05).Conclusion The use of 3D printing-assisted stent with in-vitro pre-fenestration has been proven to be safe and effective in the treatment of complex type B aortic dissection.It offers notable advantages,including the reduction of surgical duration,a lower risk of type Ⅰ internal leakage and significant redilation of the true aortic lumen after surgery.