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 relationship between oxygen consumption (VO 2), carbon dioxide production (VCO 2), and Oxygen Consumption/lactate (VO 2/Lac) with risk of death in patients with severe ARDS. Methods:A retrospective cohort study method was used, and the study subjects were hospitalized for >5 days adult patients with severe ARDS in the central intensive care unit of Henan Provincial People's Hospital from 1 March 2020 to 30 June 2023. The following patients were excluded: IC test was not completed on the 4th day of ICU admission, IC test results were unreliable, mechanical ventilation duration had exceeded 48 h at the time of ICU transfer or admission, palliative care patients and pregnant and parturient women. Using indirect calorimetry to determine VO 2 and VCO 2 values on the 4th day of admission, reviewing medical records to obtain general condition, disease information, blood gas analysis (including lactate value), diagnostic and therapeutic measures, and following up deaths by telephone and time of death. The primary outcome measure was death at 90 days, and the secondary outcome measure was death at 28 days, length of stay in ICU, total length of stay, and total hospitalization cost. Cox regression analysis and linear regression analysis were used to investigate the relationship between VO 2, VCO 2, VO 2/Lac and primary and secondary outcome indexes. Results:A total of 216 patients were enrolled, 78 patients (36.1%) died and 138 patients (63.9%) survived at 90 days. After correction for confounders, the results of multifactorial Cox regression analysis suggested that compared with the Q4 group, HR (95% CI) for 90-day risk of death in the VO 2 Q1 and Q2 groups was 3.21 (1.38, 7.49) and 3.24 (1.42, 7.38), and HR (95% CI) for 90-day risk of death in the VCO 2 Q1, Q2 and Q3 groups was 5.88 (2.33, 14.84), 4.26 (1. 60, 11.34) and 3.54 (1.34, 9.35), respectively, and the HR (95% CI) for 90-day risk of death in the VO 2/Lac Q1, Q2 and Q3 groups were 8.72 (3.01, 25.25), 8.43 (2.91, 24.47) and 4.04 (1.34, 12.17) respectively. P-trends were all <0.05, indicating that VO 2, VCO 2 and VO 2/Lac were linearly and negatively associated with the risk of 90-day mortality. In addition, VO 2, VCO 2, and VO 2/Lac were negatively associated with 28-day risk of death and higher VO 2/Lac was negatively associated with length of ICU stay. Conclusions:VO 2, VCO 2 and VO 2/Lac were negatively associated with 90-day mortality risk and 28-day mortality risk in patients with severe ARDS and may be independent risk factors predicting mortality risk of such patients.

Objective To investigate the clinical effect of simultaneous use of three endoscopes(laparoscope,choledochoscope,and duodenoscope)combined with holmium laser in the treatment of gallstones complicated with common bile duct stones.Methods A prospective randomized controlled study was carried out.A total of 80 patients with cholecystolithiasis combined with choledocholithiasis in our hospital from August 2022 to January 2024 were selected and divided into three-endoscope group(n=40)and two-endoscope group(n=40)according to random number table method.Patients in the three-endoscope group underwent laparoscopic cholecystectomy(LC)and laparoscopic common bile duct exploration(LCBDE)combined with holmium laser lithotripsy,primary closure of the common bile duct,endoscopic retrograde cholangiopancreatography(ERCP),and endoscopic nasobiliary drainage(ENBD),while the two-endoscope group underwent LC+LCBDE+T-tube drainage.The intraoperative bleeding volume,operation time,hospitalization time,hospitalization costs,and postoperative complications between the two groups were observed and analyzed.Results There was no significant difference in intraoperative bleeding volume between the two groups[(55.6±14.3)ml vs.(53.2±16.3)ml,t=0.703,P=0.484].The operation time of the three-endoscope group was significantly longer than that in the two-endoscope group[214.5(171.5,246.8)min vs.178.0(151.0,227.8)min,Z=-2.069,P=0.039].The length of hospital stay in the three-endoscope group was significantly shorter than that in the two-endoscope group[(13.2±3.3)d vs.(16.2±3.7)d,t=3.864,P=0.001].The hospitalization costs were significantly higher in the three-endoscope group than those in the two-endoscope group[39 316.0(32 338.5,43 421.0)yuan vs.33 717.0(30 873.3,37 813.3)yuan,Z=-3.272,P=0.001].There were no significant differences in the incidence of postoperative pancreatitis[2.5%(1/40)vs.2.5%(1/40),χ2=0.000,P=1.000],the incidence of bile leakage[7.5%(3/40)vs.2.5%(1/40),χ2=0.263,P=0.615],and residual stone rate[7.5%(3/40)vs.0.0%(0/40),P=0.241]between the two groups.Forty cases in the three-endoscope group were followed up for 2-20 months,with a median of 13 months.One case of stone recurrence occurred at 8 months after surgery.Forty patients in the two-endoscope group were followed up for 1-21 months,with a median of 15 months.There was 1 case of stone recurrence at 6 and 10 months postoperatively,respectively.There was no statistically significant difference in stone recurrence rate between the two groups[2.5%(1/40)vs.5.0%(2/40),χ2=0.000,P=1.000].Conclusions LC+LCBDE combined with holmium laser lithotripsy,primary closure of the common bile duct,ERCP,and ENBD in the treatment of cholecystolithiasis and choledocholithiasis can shorten the hospitalization time.The replacement of T-tube with nasobiliary duct protects the function of the Oddi sphincter,which is more in line with the concept of minimally invasive surgery and worthy of clinical application.

Objective:To screen and analyze the carcinogenesis-related differential expression profile of long non-coding RNAs(LncRNAs)in oral lichen planus(OLP)mucosa tissue,and preliminarily analyze their functions,to explore their possible role in the development of OLP.Methods:High-throughput sequencing technology was used to construct differential expression profile from 5 cases of erosive OLP lesions and 5 of normal oral mucosa.LncRNAs that are closely related to the carcinogenesis of OLP were ob-tained by bioinformatics analysis.Results:400 LncRNAs associated with OLP were screened,of which 250 were up-regulated and 150 were down-regulated,and 5 LncRNAs were obtained with differential expression associated with OLP carcinogenesis:LncRNA 54055,100128560,399717,378825 and 100130231.Conclusion:400 LncRNAs are differentially expressed in the mucosa of erosive OLP lesions,and 5 of them are related to the incidence and carcinogenesis tendency of OLP.

Objective:To screen and analyze the carcinogenesis-related differential expression profile of long non-coding RNAs(LncRNAs)in oral lichen planus(OLP)mucosa tissue,and preliminarily analyze their functions,to explore their possible role in the development of OLP.Methods:High-throughput sequencing technology was used to construct differential expression profile from 5 cases of erosive OLP lesions and 5 of normal oral mucosa.LncRNAs that are closely related to the carcinogenesis of OLP were ob-tained by bioinformatics analysis.Results:400 LncRNAs associated with OLP were screened,of which 250 were up-regulated and 150 were down-regulated,and 5 LncRNAs were obtained with differential expression associated with OLP carcinogenesis:LncRNA 54055,100128560,399717,378825 and 100130231.Conclusion:400 LncRNAs are differentially expressed in the mucosa of erosive OLP lesions,and 5 of them are related to the incidence and carcinogenesis tendency of OLP.

Objective To investigate the clinical effect of simultaneous use of three endoscopes(laparoscope,choledochoscope,and duodenoscope)combined with holmium laser in the treatment of gallstones complicated with common bile duct stones.Methods A prospective randomized controlled study was carried out.A total of 80 patients with cholecystolithiasis combined with choledocholithiasis in our hospital from August 2022 to January 2024 were selected and divided into three-endoscope group(n=40)and two-endoscope group(n=40)according to random number table method.Patients in the three-endoscope group underwent laparoscopic cholecystectomy(LC)and laparoscopic common bile duct exploration(LCBDE)combined with holmium laser lithotripsy,primary closure of the common bile duct,endoscopic retrograde cholangiopancreatography(ERCP),and endoscopic nasobiliary drainage(ENBD),while the two-endoscope group underwent LC+LCBDE+T-tube drainage.The intraoperative bleeding volume,operation time,hospitalization time,hospitalization costs,and postoperative complications between the two groups were observed and analyzed.Results There was no significant difference in intraoperative bleeding volume between the two groups[(55.6±14.3)ml vs.(53.2±16.3)ml,t=0.703,P=0.484].The operation time of the three-endoscope group was significantly longer than that in the two-endoscope group[214.5(171.5,246.8)min vs.178.0(151.0,227.8)min,Z=-2.069,P=0.039].The length of hospital stay in the three-endoscope group was significantly shorter than that in the two-endoscope group[(13.2±3.3)d vs.(16.2±3.7)d,t=3.864,P=0.001].The hospitalization costs were significantly higher in the three-endoscope group than those in the two-endoscope group[39 316.0(32 338.5,43 421.0)yuan vs.33 717.0(30 873.3,37 813.3)yuan,Z=-3.272,P=0.001].There were no significant differences in the incidence of postoperative pancreatitis[2.5%(1/40)vs.2.5%(1/40),χ2=0.000,P=1.000],the incidence of bile leakage[7.5%(3/40)vs.2.5%(1/40),χ2=0.263,P=0.615],and residual stone rate[7.5%(3/40)vs.0.0%(0/40),P=0.241]between the two groups.Forty cases in the three-endoscope group were followed up for 2-20 months,with a median of 13 months.One case of stone recurrence occurred at 8 months after surgery.Forty patients in the two-endoscope group were followed up for 1-21 months,with a median of 15 months.There was 1 case of stone recurrence at 6 and 10 months postoperatively,respectively.There was no statistically significant difference in stone recurrence rate between the two groups[2.5%(1/40)vs.5.0%(2/40),χ2=0.000,P=1.000].Conclusions LC+LCBDE combined with holmium laser lithotripsy,primary closure of the common bile duct,ERCP,and ENBD in the treatment of cholecystolithiasis and choledocholithiasis can shorten the hospitalization time.The replacement of T-tube with nasobiliary duct protects the function of the Oddi sphincter,which is more in line with the concept of minimally invasive surgery and worthy of clinical application.

Nonalcoholic fatty liver disease （NAFLD） is an abnormal lipid metabolic disorder of the liver characterized by accumulation of a large amount of lipids in the liver， and it is currently the most common liver disease around the world. Mendelian randomization （MR） incorporates genomic data into traditional epidemiological study designs to infer the causal relationship between exposure factors and disease risk. In recent years， MR has been widely used in studies on inference of the etiology of NAFLD. This article systematically summarizes the advances in the application of MR in NAFLD research， so as to provide new ideas for understanding the nature of the disease and scientific interventions.

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.

Humans ; Hypertension, Pulmonary/surgery* ; Learning Curve ; Angioplasty, Balloon ; Pulmonary Embolism/therapy* ; Chronic Disease ; Pulmonary Artery/surgery*