Objective To investigate the impact of long non-coding RNA maternal expression gene 3(lncRNA MEG3)on hypoxia/reoxygenation(H/R)-induced cardiomyocyte apoptosis by modulating the miR-202-5p/signal transducer and activator of transcription 3(STAT3)axis.Methods An H/R cell model was constructed and randomly separated into four groups:sh-NC(transfected with NC shRNA),sh-MEG3(transfected with lncRNA MEG3 shRNA),miR-NC(transfected with lncRNA MEG3 shRNA and NC miR),and in-miR-202-5p(transfected with lncRNA MEG3 shRNA and miR-202-5p inhibitor).In addition,the H/R group(nontransfected H/R model cells)and the AC 16 group(normal AC 16 cells)were set.Quantitative real-time PCR method was used to analyze the expression of lncRNA MEG3,miR-202-5p,and STAT3 in cells from each group.CCK-8 method was used to analyze cell viability.Flow cytometry was used to analyze apoptosis.Enzyme-linked immunosorbent,colorimetric,and probe assays were applied to detect the levels of inter-leukin-6(IL-6),tumor necrosis factor α(TNF-α),malondialdehyde(MD A),glutathione peroxidases(GSH-Px),and reactive oxygen spe-cies(ROS).Western blotting was carried out to examine the expression of STAT3,Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),caspase-3,and caspase-9.A dual luciferase assay was used to analyze the relationship between lncRNA MEG3 and miR-202-5p,as well as the relationship between miR-202-5p and STAT3.Results Compared with that in the AC 16 group,the expression of lncRNA MEG3 and STAT3 in cells in the H/R group increased,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the expression of lncRNA MEG3 and STAT3 decreased in the sh-MEG3 group,while the expression of miR-202-5p increased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the expression of lncRNA MEG3 and STAT3 increased in the in-miR-202-5p group,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the AC 16 group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the H/R group.In contrast,the cell viability,clone count,levels of superoxide dismutase(SOD)and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 increased in the sh-MEG3 group.In contrast,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 decreased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the in-miR-202-5p group.In contrast,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).There were multiple binding sites between lncRNA MEG3 and miR-202-5p,and between miR-202-5p and STAT3.The luciferase activity was lower in the WT-MEG3+miR-202-5p group than in the WT-MEG3+miR-NC group(P＜0.05).The luciferase activity was lower in the WT-STAT3+miR-202-5p group than in the WT-STAT3+miR-NC group(P＜0.05).Conclusion Knocking down lncRNA MEG3 can downregulate STAT3 by negatively regulating miR-202-5p,inhibiting H/R-induced cardiomyocyte apoptosis.

Objective To investigate the impact of long non-coding RNA maternal expression gene 3(lncRNA MEG3)on hypoxia/reoxygenation(H/R)-induced cardiomyocyte apoptosis by modulating the miR-202-5p/signal transducer and activator of transcription 3(STAT3)axis.Methods An H/R cell model was constructed and randomly separated into four groups:sh-NC(transfected with NC shRNA),sh-MEG3(transfected with lncRNA MEG3 shRNA),miR-NC(transfected with lncRNA MEG3 shRNA and NC miR),and in-miR-202-5p(transfected with lncRNA MEG3 shRNA and miR-202-5p inhibitor).In addition,the H/R group(nontransfected H/R model cells)and the AC 16 group(normal AC 16 cells)were set.Quantitative real-time PCR method was used to analyze the expression of lncRNA MEG3,miR-202-5p,and STAT3 in cells from each group.CCK-8 method was used to analyze cell viability.Flow cytometry was used to analyze apoptosis.Enzyme-linked immunosorbent,colorimetric,and probe assays were applied to detect the levels of inter-leukin-6(IL-6),tumor necrosis factor α(TNF-α),malondialdehyde(MD A),glutathione peroxidases(GSH-Px),and reactive oxygen spe-cies(ROS).Western blotting was carried out to examine the expression of STAT3,Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),caspase-3,and caspase-9.A dual luciferase assay was used to analyze the relationship between lncRNA MEG3 and miR-202-5p,as well as the relationship between miR-202-5p and STAT3.Results Compared with that in the AC 16 group,the expression of lncRNA MEG3 and STAT3 in cells in the H/R group increased,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the expression of lncRNA MEG3 and STAT3 decreased in the sh-MEG3 group,while the expression of miR-202-5p increased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the expression of lncRNA MEG3 and STAT3 increased in the in-miR-202-5p group,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the AC 16 group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the H/R group.In contrast,the cell viability,clone count,levels of superoxide dismutase(SOD)and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 increased in the sh-MEG3 group.In contrast,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 decreased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the in-miR-202-5p group.In contrast,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).There were multiple binding sites between lncRNA MEG3 and miR-202-5p,and between miR-202-5p and STAT3.The luciferase activity was lower in the WT-MEG3+miR-202-5p group than in the WT-MEG3+miR-NC group(P＜0.05).The luciferase activity was lower in the WT-STAT3+miR-202-5p group than in the WT-STAT3+miR-NC group(P＜0.05).Conclusion Knocking down lncRNA MEG3 can downregulate STAT3 by negatively regulating miR-202-5p,inhibiting H/R-induced cardiomyocyte apoptosis.

Necroptosis is a regulated process of programmed cell death independent of aspartic acid-specific cysteine protease,which can induce inflammation.Studies have shown that necroptosis is closely related to the progression and prognosis of pancreatic disease and plays an important two-way regulatory role in its progression.Related necroptosis inhibitors and inducers are expected to be used in the treatment of pancreatic disease.We herein review the mechanism of necroptosis and its role in the progression of pancreatic disease to provide a new understanding of the pathogenesis and treatment of pancreatic diseases and offer a theoretical basis for the research and development of targeted drugs.

ObjectiveThis study aims to examine the effect of Rhei Radix et Rhizoma-Coptidis Rhizoma on reducing insulin resistance in db/db mice by regulating the adenylate activated protein kinase （AMPK）/UNC-51-like kinase 1 （ULK1）/key molecule of autophagy， benzyl chloride 1 （Beclin1） pathway and elucidate the underlying mechanism. MethodSixty 6-week-old male db/db mice were studied. They were randomly divided into the model group， metformin group （0.26 g·kg^-1）， and low-， middle-， and high-dose groups （2.25， 4.5， 9 g·kg^-1） of Rhei Radix et Rhizoma-Coptidis Rhizoma. A blank group of db/m mice of the same age was set， with 12 mice in each group. After eight weeks of continuous intragastric administration， the blank group and model group received distilled water intragastrically once a day. The survival status of the mice was observed， and fasting blood glucose （FBG） was measured using a Roche blood glucose device. Fasting serum insulin （FINS） was measured using an enzyme-linked immunosorbent assay， and the insulin resistance index （HOMA-IR） was calculated. Hematoxylin-eosin （HE） staining was performed to observe the pathological changes in the liver of the mice. The protein expression levels of AMPK， Beclin1， autophagy associated protein 5 （Atg5）， and p62 in liver tissue were determined by using Western blot. The protein expression levels of autophagy associated protein 1 light chain 3B （LC3B） and ULK1 in liver tissue were determined using immunofluorescence. Real-time fluorescence quantitative PCR （Real-time PCR） was used to measure mRNA expression levels of AMPK， Beclin1， Atg5， ULK1， and p62. ResultCompared with the blank group， the model group exhibited a significant increase in body mass （P<0.01）. Additionally， the levels of FBG， FINS， and HOMA-IR significantly changed （P<0.01）. The structure of liver cells was disordered. The protein expression levels of AMPK， Beclin1， and Atg5 in liver tissue were significantly decreased （P<0.01）， while the expression level of p62 protein was significantly increased （P<0.01）. The expression levels of mRNA and proteins were consistent. Compared with the model group， the body mass of the metformin group and high and medium-dose groups of Rhei Radix et Rhizoma-Coptidis Rhizoma was significantly decreased （P<0.05）. FBG， FINS， and HOMA-IR were significantly decreased （P<0.05，P<0.01）. After treatment， the liver structure damage in each group was alleviated to varying degrees. The protein expressions of AMPK， Beclin1， Atg5， LC3B， and ULK1 were increased （P<0.05，P<0.01）， while the protein expression of p62 was decreased （P<0.01）. The expression levels of mRNA and proteins were generally consistent. ConclusionThe combination of Rhei Radix et Rhizoma-Coptidis Rhizoma can effectively improve liver insulin resistance， regulate the AMPK autophagy signaling pathway， alleviate insulin resistance in db/db mice， and effectively prevent the occurrence and development of type 2 diabetes.

ObjectiveTo analyze the clinical characteristics and the use of traditional Chinese medicine （TCM） therapy in cancer patients with COVID-19， and to provide reference for making TCM prevention and treatment strategies and determining diagnosis and treatment priorities for patients with malignant tumors in the COVID-19 epidemic. MethodsThe medical records of 225 malignant tumor cases with COVID-19 who were admitted to 7 national research centers from January 1st to 20th， 2023 were retrospectively collected， and the main symptoms and duration after infection， nucleic acid negative conversion time， use of TCM therapy， and changes in adverse reactions after resuming anti-tumor treatment were analyzed. ResultsA total of 222 malignant tumor patients with COVID-19 were included in the analysis， involving 205 mild cases and 17 moderate cases. The top four most frequently reported clinical symptoms were fever （165 cases）， expectoration or dry cough （99 cases）， decreased appetite （95 cases） and fatigue （85 cases）， of which 40 expectoration or dry cough cases ， 37 fatigue cases and 29 decreased appetite cases lasted for more than 14 days. One hundred and five patients with nucleic acid detection report had a median negative conversion time of 14 days. The nucleic acid negative conversion time was significantly longer in patients with lung cancer compared to those with digestive system malignant tumors， and in those with myelosuppression than those without （P＜0.01）. During the infection period， 47.30% （105/222） of the patients used Chinese patent medicine， and 21.17% （47/222） were treated with herbal decoctions. The use of TCM in patients during the prevention and rehabilitation period， was 1.80%（4/222） and 7.21%（16/222）， respectively. Fifty-five patients resumed anti-tumor treatment after nucleic acid negative conversion， and received TCM simultaneously. Observed adverse reactions， including gastrointestinal reactions， bone marrow suppression， and neurotoxicity， were all grade 1 to 2， and no new adverse events occurred during follow-up. ConclusionCertain malignant tumor patients with COVID-19 had prolonged symptoms and nucleic acid negative conversion time Rational use of TCM can help to promote the rehabilitation of the patients and ensure the smooth process of anti-tumor treatment after infection.

Peptide-based hydrogel, the polymer materials with a special network structure, are widely used in various fields of biomedicine due to their stable properties and biocompatibility. Environment-responsive self-assembled peptide aqueous solutions can respond to environment changes by the self-assembly of peptides into nanofiber networks. Peptide-based hydrogels well simulate the extracellular matrix and cell growth microenvironment, being suitable for 3D cell culture and organoid culture. To establish a tumor organoid culture system with peptide-based hydrogels, we cultured Panc-1, U87, and H358 cells in a 3D spherical manner using CulX Ⅱ peptide-based hydrogels in 24-well plates for 15 days. The organoids showed a 3D spherical shape, and their sizes increased with the extension of the culture time, with a final diameter ranging from 150 to 300 μm. The organoids had a large number, varying sizes, good cell viability, clear edges, and a good shape, which indicated successful organoid construction. The tumor organoid culture system established in this study with CulX Ⅱ peptide-based hydrogels provides a model for studying tumor pathogenesis, drug development, and tumor suppression.
Hydrogels/chemistry* ; Organoids ; Humans ; Peptides/pharmacology* ; Cell Line, Tumor ; Cell Culture Techniques, Three Dimensional ; Cell Culture Techniques ; Cell Survival/drug effects* ; Nanofibers/chemistry*

ObjectiveTo reveal the intervention effect of Dahuang Mudantang on pancreatic injury in rats with acute pancreatitis （AP） of dampness-heat in large intestine syndrome and explore its possible mechanism based on network pharmacology. MethodNinety-six SPF-grade Wistar rats were randomly divided into the following six groups： a blank group， a model group， low-， medium-， and high-dose Dahuang Mudantang groups （3.5， 7， and 14 g·kg^-1）， and a Qingyi Lidan granules group （3 g·kg^-1）， with 16 rats in each group. The AP model of dampness-heat in large intestine syndrome was induced in rats except for those in the blank group by "high-temperature and high-humidity environment + high-sugar and high-fat diet + retrograde injection of 5% sodium taurocholate into the pancreaticobiliary duct". The blank and model groups received equal volumes of distilled water by gavage， while the treatment groups were administered Dahuang Mudantang or Qingyi Lidan granules 1 hour before modeling， and 12 and 24 hours after modeling. Samples were collected 1 hour after the last administration. The general conditions of the rats were observed. The AP model of dampness-heat in large intestine syndrome was evaluated. Serum amylase （AMS） and C-reactive protein （CRP） levels were determined using biochemical methods. Pancreatic tissue morphology was observed using hematoxylin-eosin （HE） staining. Network pharmacology was employed to predict potential targets of Dahuang Mudantang in the intervention in AP， and molecular biology technique was used to verify relevant targets. ResultCompared with the blank group， the model group exhibited lethargy， unkempt fur， loose and foul-smelling stools， elevated anal temperature with arching and twisting reactions， significantly increased serum levels of AMS and CRP （P<0.05）， abnormal pancreatic ductules， disordered interlobular spaces， and inflammatory cell infiltration in histopathological examination， as well as pathological changes including pancreatic acinar cell swelling， congestion， and necrosis. Compared with the model group， the treatment groups showed varying degrees of improvement in general survival conditions， reduced twisting reactions， visibly improved stool characteristics， reduced pancreatic tissue edema and necrosis， decreased serum AMS and CRP levels （P<0.05）， with the high-dose Dahuang Mudantang group showing the most pronounced effects （P<0.05）. Network pharmacology prediction indicated that hederagenin， β-sitosterol， and quercetin were the most widely connected active compounds with disease targets. Protein-protein interaction （PPI） network analysis revealed that protein kinase B （Akt）， tumor protein P53 （TP53）， tumor necrosis factor （TNF）， interleukin-6 （IL-6）， transcription factor （JUN）， vascular endothelial growth factor α （VEGFα）， interleukin-1β （IL-1β）， and vascular cell adhesion molecule-1 （VCAM1） were key targets in the "drug-disease" interaction. KEGG enrichment analysis suggested that the response of the mitogen activated protein kinase （MAPK） signaling pathway might be a core mechanism for DHMDT in the intervention in AP. Molecular biology analysis showed that compared with the blank group， the model group had significantly increased levels of TNF-α， IL-6， and VCAM-1 in pancreatic tissue （P<0.05）， as well as significantly elevated expression levels of p38 mitogen-activated protein kinase （p38 MAPK）， mitogen-activated protein kinase-activated protein kinase 2 （MK2）， and human antigen R （HUR） genes and proteins （P<0.05）. Compared with the model group， the treatment groups exhibited decreased levels of TNF-α， IL-6， and VCAM-1 in pancreatic tissue （P<0.05）， reduced expression levels of p38 MAPK， MK2， and HUR genes and proteins， with the high-dose Dahuang Mudantang group showing the most pronounced effects （P<0.05）. ConclusionDahuang Mudantang activates and regulates the p38 MAPK/MK2/HUR signaling pathway to suppress the release of inflammatory factors， thereby improving pancreatic injury.

ObjectiveTo explore the mechanism of Dahuang Mudantang in alleviating the intestinal injury in the rat model of acute pancreatitis via the high-mobility group box 1 （HMGB1）/receptor for advanced glycation endproduct （RAGE）/nuclear factor-κB （NF-κB） signaling pathway. MethodOne hundred and twenty SPF-grade Wistar rats received retrograde injection of 5% sodium taurocholate into the biliopancreatic duct for the modeling of intestinal injury in acute pancreatitis. The rats were randomized into blank， model， low-， medium-， and high-dose （3.5， 7， 14 g·kg^-1， administrated by gavage） Dahuang Mudantang， and octreotide （1×10^-5 g·kg^-1， subcutaneous injection） groups （n=20）. The rats in blank and model groups received equal volume of distilled water by gavage. Drugs were administered 1 h before and every 12 h after modeling， and samples were collected 24 h after modeling. The general status of the rats was observed. The biochemical methods were employed to measure the levels of amylase （AMS） and C-reactive protein （CRP） in the serum. The enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6 in the colon tissue. The morphological changes of pancreatic and colon tissues were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were employed to measure the expression levels of HMGB1， RAGE， inhibitor of NF-κB kinase （IKK）， and NF-κB suppressor protein α（IκBα）in the colon tissue. ResultThe rats in the model group showed poor general survival， writhing response， reduced frequency of defecation， and dry stool. The symptoms of rats in the model group were mitigated in each treatment group， and the high-dose Dahuang Mudantang showed the most significant effect. Compared with the normal group， the model group had elevated AMS and CRP levels （P<0.05）， which were lowered by Dahuang Mudantang （P<0.05）， especially that at the high dose （P<0.05）. Compared with the normal group， the modeling elevated that levels of TNF-α， IL-1β， and IL-6 （P<0.05）. Such elevations were lowered by Dahuang Mudantang （P<0.05）， and the high-dose group and the octreotide group showed better performance （P<0.05）. The modeling caused necrotic， congested， and destructed pancreatic and colonic tissues， which were ameliorated by the drugs， especially high-dose Dahuang Mudantang. Compared with the normal group， the modeling up-regulated the mRNA levels of HMGB1， RAGE， IKK， IκBα， and NF-κB （P<0.05）. Compared with the model group， Dahuang Mudantang and octreotide down-regulated the mRNA levels of HMGB1， RAGE， IKK， IκBα， and NF-κB （P<0.05）， and the high-dose Dahuang Mudantang demonstrated the best performance （P<0.05）. Western blot results showed a trend consistent with the results of Real-time PCR. ConclusionDahuang Mudantang can improved the general status， reduce inflammation， and alleviate histopathological changes in the pancreatic and colon tissues in the rat model of acute pancreatitis by inhibiting the HMGB1/RAGE/NF-κB signaling pathway.

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis and neuromyelitis optica spectrum disorders (NMOSD) are central nervous system diseases mediated by autoimmune antibodies. With the improvement of diagnosis and treatment, the reports of the two diseases appearing in the same patient are increasing. To strengthen the understanding of this kind of comorbidity, two cases of anti-NMDAR-encephalitis coexisting with NMOSD admitted in our hospital were reported. The clinical manifestations and imaging examination of the two patients showed the evidence of involvement of both brain and spinal cord. The anti-NMDAR antibody and anti-aquaporin 4 antibody tests were positive. In addition, gamma globulin or corticosteroid impulse therapy was effective in the treatment of two patients.

Objective The purpose of this study was to explore common complications and their clinicopathological features in pediatric liver transplantation.Methods Clinical and pathological data of 240 liver biopsies from 168 children that conducted liver puncture from January 2015 to May 2018 in Tianjin First Central Hospital was retrospectively analyzed.We comprehensively analyzed incidence rate and pathological features of various complications,and correlations between acute rejection and C4d staining result or Banff score.Results A total of 86.67％ (208/240) liver biopsies could be definitely diagnosed with incidence rate of main complications in descending order as follows:T cell mediated rejection (TCMR) 60.57％ (126/208),drug-induced liver injury (DILI) 17.31％ (36/208),biliary complication 8.17％ (17/208),vascular complication 3.37％ (7/208),ischemia/reperfusion injury (IRI) 2.88％ (6/208),antibody mediated acute rejection (AMR) 1.92％ (4/208),HBV infection 1.92％ (4/208),non-alcoholic fatty liver disease (NAFLD) 1.44％ (3/208),chronic rejection (CR) 0.96 ％ (2/208) and HCV infection 0.48 ％ (1/208).TCMR and AMR in acute rejection (AR) accounted for 96.92％ (126/130) and 3.08％ (4/160),and into(portal-based,PB)type TCMR accounted for 96.03％(121/126) with the detectable rate of BP type subtype TCMR of 26.45％(32/121)within 30 d.There were 65.87％ (83/126)、25.40％ (32/126) 和4.76％ (6/126) of BP TCMR samples with "Banff ACR RAI" score within 3-5,6-7 and 8-9,and RAI score was negatively correlated with postoperative time (r =0.127,P =0.084).The incidence rate of central perivenulitis (CP) and portal eosinophils infiltration (PEI) in BP TCMR was 63.63％ (77/121) 和43.80％ (53/ 121),respectively,additionally,the PEI level was positively correlate with RAI score (P＜0.05).CP TCMR and AMR occurred within 30d-365 d and 8 d-180 d,respectively postoperative,while,the two CR occurred at 1095 d and 1335 d postoperative,and significant correlation was strikingly observed between rejection subtype and postoperative time (Z =9.231,P =0.026).C4d positive rate was 10％ (24/240),which was associated with Banff score and postoperative time,besides,C4d score was also correlated with rejection subtype and RAI score.The occurrence of DILI was mainly at time of ＜90 d or ＞180 d postoperative,and the detectable rate of biliary complication within 180 d postoperative was 82.35％ (14/17),IRI Appear in ＜30d.Hepatic artery complication account for nearly 57.14％ (4/7),occurrence time is ≤90 d.Occurrence of HBV infection,CMV infection and NAFLD were mainly at ＞365 d,＜90 d and ＜365 d,respectively.Conclusion There were lots of differences in clinical and pathological features among multi pediatric liver transplantation complications.Liver puncture plays an important role in rejection subtype classification and grading,as well as in non-rejection complications identification.