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 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 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 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.

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 of individualized coil embolization in the interventional treatment of renal artery aneurysm(RAA).Methods Data of 23 patients with RAA treated by individualized coil embolization were analyzed.There were 27 RAAs,in which narrow-necked RAAs were treated with coil embolization and wide-necked RAAs were treated with stent-assisted coil embolization.The efficacy of the two embolization methods were analyzed and the changes of renal function and symptoms were observed.Results A total of 27 RAAs in 23 patients were successfully embolized at one time,including 23 narrow-necked RAAs in 19 cases treated with coil embolization and 4 wide-necked RAAs in 4 patients treated with stent-assisted coil embolization.The embolization effect of 20 cases(86.96%)reached Raymond grade Ⅰ,and 3 cases(13.04%)reached gradeⅡ.Postoperative computed tomography angiography(CTA)showed that all parent arteries were patent,the RAA was not visualized,and there was no renal infarction.There was no statistical difference in creatinine values before operation,1 month,6 months and 1 year after operation(P>0.05).In the 12 patients with hypertension,there were statistically significant differences in blood pressure at 1 year after operation compared with preoperative,1 month,and 6 months after operation(P<0.05).The symptoms of low back pain and hematuria disappeared after operation.Conclusion Individualized coil embolization for RAA is safe,effective and worthy of clinical promotion.

OBJECTIVE To investigate the inhibitory effect and mechanism of lead(Pb2+)on γ-amino-butyric acid(GABA)A receptor-mediated currents(IGABA)and GABAergic synaptic transmission in rat cortical neurons.METHODS ①The cortical neurons from 0 d Sprague Dawley(SD)rats were cultured for experiments.The cultured cells(7-14 d)were recorded using the patch-clamp technique to analyze the effects of Pb2+ at different concentrations(1,5,10,50 and 100 μmol·L-1)on IGABA induced by GABA 100 μmol·L-1.② The effects of Pb2+ 50 μmol·L-1 on IGABA induced by GABA at different concentrations(1,10,50,100,500 and 100 μmol·L-1)were detected.③Brain slices(350 μm)were prepared from SD rats(15-19 d).The spontaneous inhibitory post-synaptic currents(sIPSCs),miniature inhibitory post-synaptic currents(mIPSCs)and current injection-induced action potential(AP)were recorded to detect the effects of Pb2+ 10 μmol·L-1 on the amplitude and frequency of sIPSCs and mIPSCs,and the frequency of AP.RESULTS ①Pb2+ inhibited IGABA in a concentration-dependent manner,and IC50 was(68±20)μmol·L-1.②Pb2+ also suppressed the maximum current induced by GABA(P<0.01),with a significant increase of the GABA′s EC50 from(20±6)μmol·L-1 to(87±39)μmol·L-1,indicating that Pb2+ might inhibit IGABA in a non-competitive mechanism.③Pb2+ 10 μmol·L-1 inhibited the frequency(P<0.01)rather than the ampli-tude of sIPSCs reversibly,but had no effect on eigher the frequency or amplitude of mIPSCs.In addi-tion,Pb2+ decreased the frequency of evoked AP by current injection(P<0.01)and reduced the overall excitability of rat cortical neurons.CONCLUSION Pb2+ can significantly inhibit IGABA in primary cultured neurons.In the brain slice experiment,Pb2+ may affect sIPSCs frequency by inhibiting the AP of cortical neurons,suggesting that there are different intrinsic mechanisms through which Pb2+ inhibits both IGABA in primary cultured neurons and the frequency of sIPSCs in brain slice neurons,which points to the complexity of the mechanism of Pb2+ poisoning.