Objective To explore the effects and mechanisms of human umbilical cord mesenchymal stem cell (HUC-MSC)-derived exosomes (Exo) pretreated with Huangkui capsules on renal ischemia-reperfusion injury (IRI). Methods HUC-MSCs were cultured in media containing different concentrations of Huangkui capsules for 24 hours to determine cell viability and select an appropriate concentration for subsequent experiments. HUC-MSCs were pretreated with 50 μg/mL Huangkui capsules for 24 hours, and Exo were extracted using an exosome extraction kit. The morphology was observed under a transmission electron microscope, particle size was measured by nanoparticle tracking analysis, and the expression of exosomal membrane surface marker proteins was detected by Western blot. Human renal tubular epithelial cells (HK-2 cells) were randomly divided into hypoxia/reoxygenation group (M group), hypoxia/reoxygenation + Exo group (E group), and hypoxia/reoxygenation + Huangkui capsules pretreated Exo group (H group). Western blotting was used to measure the expression of endoplasmic reticulum stress (ERS)-related proteins, and real-time fluorescent quantitative reverse transcription polymerase chain reaction was used to measure the expression of ERS-related gene messenger RNA (mRNA). Mice were randomly divided into sham operation group (Sham group), ischemia-reperfusion group (I/R group), ischemia-reperfusion + Exo group (E group), and ischemia-reperfusion + Huangkui capsules pretreated Exo group (H group). Renal histological assessment, serum creatinine (Scr), blood urea nitrogen (BUN) measurement and inflammatory factor detection were performed 24 hours later. Results Both Exo and Huangkui capsules prereated Exo had a bilayer membrane structure and a cup-shaped morphology; their average particle sizes were 116.8 nm and 81.3 nm, respectively. Both expressed CD9, CD63, TSG101. Compared with the M group, the E group had decreased relative expression of transcription factor 6 (ATF6) and protein kinase R-like endoplasmic reticulum kinase (PERK) proteins, increased mRNA relative expression, increased relative expression of C/EBP homologous protein (CHOP) protein, and decreased mRNA relative expression. Compared with the E group, the H group had decreased relative expression of ATF6, PERK, CHOP proteins, and decreased mRNA relative expression of ATF6 and PERK (all P<0.05). Animal experimental results showed that compared with the Sham group, the I/R group had increased renal tubular injury scores, Scr, BUN, interleukin (IL)-1β, IL-10, IL-18, tumor necrosis factor (TNF)-α levels. Compared with the I/R group, the E and H groups had decreased renal tubular injury scores and Scr, BUN, IL-1β, IL-10, IL-18, TNF-α levels. Compared with the E group, the H group had decreased renal tubular injury scores and Scr, BUN, IL-1β, IL-10, IL-18, TNF-α levels (all P<0.05). Conclusions Huangkui capsules pretreatment HUC-MSC-derived Exo may alleviate renal IRI by inhibiting ERS.

Purpose To investigate the cell components in different tumor lineages of multiple synchronous pituitary neuro-endocrine tumors(PitNETs)/neuroendocrine tumors(MSPs)and to carry out accurate histological typing,which provides an important basis for determining the follow-up plan and adjuvant therapy after surgery.Methods The clinical data of 855 pa-tients with PitNETs were collected and reclassified according to the new WHO standard.The clinicopathological features of 13 patients diagnosed as MSPs were analyzed retrospectively.The immunohistochemical EnVision two-step method was used to de-tect the expression of PIT-1,SF-1,T-PIT,GH,PRL,TSH,LH,FSH,ACTH,etc.,and related literatures were reviewed.Methods A total of 855 cases of pituitary neuroendocrine tumor from the second hospital of the Chinese Hebei Medical U-niversity were collected and reclassified according to the new WHO standard,and review the literature.Results(1)The age of patients were 39-68 years with median age of 55 years.7 cases were female and 6 were male;(2)Imaging findings:There was 1.2～3.8 cm(mean 2.5 cm)in maximum tumor di-ameter.13 patients were large adenomas,neither MSPs nor sin-gle lineage PitNETs could not assessed on imaging;(3)Clinical manifestations:3 cases had hyperprolactinemia which all contai-ning PitNETs components of PRL,one case was immature PIT-1 polyhormone cell tumor,one was dense granular prolactinoma,and one case was eosinophilic stem tumor.One patient had Cushing's disease and contained a Crook cell tumor component;two had elevated ACTH,and one had an adrenocorticotropic ad-enomatous component.In the two patients with no evidence of hormone excess,all contained gonadotropin cell tumor compo-nents;(4)Combination form:11 cases of the combination of the two cell lineages(5 cases of combination of SF-1 lineage and PIT-1 lineage;4 cases of combination of T-PIT lineage and PIT-1 lineage;1 case of null cell tumor and PIT-1 lineage;1 case of plurihormonal PitNETs and SF-1 lineage);Two cases of three cell combinations(null cell tumor,PIT-1 lineage,and T-PIT lineage);Among them,13 cases were PIT-1 lineage tumors,46.2％(6/13)were gonadotropin cell tumor,38.5％(5/13)were prolactinoma;(5)The presence of high-risk lineage tumors in the 10 patient combinations:3 immature PIT-1-lineage tumor,1 Crooke cell PitNETs,1 acidophil stem cells tumor,3 zero-cell PitNETs,and 4 silent-type sparse granular adrenocorti-cotropic hormone PitNETs;Two of them were combinations of two high-risk subtypes.Conclusion MSPs in our center are large adenomas,although their incidence is only 1.5％of Pit-NETs,2/3 cases have high-risk lineage tumor components,and the use of pituitary cell lineage transcription factors and adeno-hypophyseal hormones plays an important role in distinguishing and clarifying the different components of MSPs.

The typical hallmark of tumor evolution is metabolic dysregulation. In addition to secreting immunoregulatory metabolites, tumor cells and various immune cells display different metabolic pathways and plasticity. Harnessing the metabolic differences to reduce the tumor and immunosuppressive cells while enhancing the activity of positive immunoregulatory cells is a promising strategy. We develop a nanoplatform (CLCeMOF) based on cerium metal-organic framework (CeMOF) by lactate oxidase (LOX) modification and glutaminase inhibitor (CB839) loading. The cascade catalytic reactions induced by CLCeMOF generate reactive oxygen species "storm" to elicit immune responses. Meanwhile, LOX-mediated metabolite lactate exhaustion relieves the immunosuppressive tumor microenvironment, preparing the ground for intracellular regulation. Most noticeably, the immunometabolic checkpoint blockade therapy, as a result of glutamine antagonism, is exploited for overall cell mobilization. It is found that CLCeMOF inhibited glutamine metabolism-dependent cells (tumor cells, immunosuppressive cells, etc.), increased infiltration of dendritic cells, and especially reprogrammed CD8⁺ T lymphocytes with considerable metabolic flexibility toward a highly activated, long-lived, and memory-like phenotype. Such an idea intervenes both metabolite (lactate) and cellular metabolic pathway, which essentially alters overall cell fates toward the desired situation. Collectively, the metabolic intervention strategy is bound to break the evolutionary adaptability of tumors for reinforced immunotherapy.

Objective To explore the extraction and purification method of Kupffer and hepatic stellate cells from mouse liver and provide references and suggestions for the separation and extraction method ology of primary non-parenchymal cells from mouse liver.Methods After in vivo collagenase perfusion digestion,various reagents and method,such as Percoll and OptiPrep,were used to extract C57BL/6 mice Kupffer and hepatic stellate cells,and evaluate their purity by flow cytometry and immunofluorescence.Results The two-layer Percoll method to extract Kupffer cells and the two-layer OptiPrep method to extract hepatic stellate cells were feasible,and purity reached>90%.The cell yield was 1～2×107/liver,and the cell survival rate was>90%.After 48 hours of primary cell culture,the number of F4/80-positive Kupffer cells and α-SMA-positive hepatic stellate cells reached>90%.Conclusions The separation and extraction method of Kupffer and hepatic stellate cells from mouse liver are perfect,reliable,cost-effective,and reproducible.

Objective:To investigate the effect of autologous hematopoietic stem cell transplantation (auto-HSCT) bridging to chimeric antigen receptor T cell (CAR-T) immunotherapy for follicular lymphoma (FL) transformed to B-lymphoblastic leukemia/lymphoma (B-LBL).Methods:The diagnosis and treatment of 1 patient with FL transformed to B-LBL admitted to the First Hospital of Soochow University in August 2020 were retrospectively analyzed and the literature was reviewed.Results:The male patient was 65 years old, and was diagnosed as FL (stage Ⅳ group A, FL international prognostic index -1 score 3 points, high-risk group) in August 2020. And then he was given 6 courses of RB (rituximab combined with bendamustine) regimen, with complete remission (CR) at mid-term and end-stage PET-CT, followed by regular maintenance therapy with rituximab every 2 months, and disease progressed after 4 courses of maintenance therapy. According to the results of histopathology and bone marrow aspiration in December 2021, he was diagnosed B-LBL involving the bone marrow. Partial remission was achieved after induction therapy with zanubnulindb combined with hyper CVAD (cyclophosphamide + doxorubicin + vindesine + dexamethasone) regimen, followed by auto-HSCT bridging to CAR-T treatment targeting CD19 and CD22, which proceeded smoothly with cytokine release syndrome grade 0, immune effector cell-associated neurotoxicity syndrome grade 0. The patient successfully underwent hematopoietic reconstruction and orally taken ibrutinib after discharge. PET-CT indicated CR 2 months after transplantation and he was still in disease-free survival state.Conclusions:The prognosis of FL transformed to B-LBL is poor, and auto-HSCT bridging to CAR-T can improve the prognosis and prolong the survival time of patients who cannot undergo allogeneic hematopoietic stem cell transplantation.

Objective The intervention effect of Huoxue Jiedu Huayu Recipe on the pyroptosis of vascular smooth muscle cells(VSMC)on the contralateral renal vessel of 6-month unilateral ureteral obstruction(UUO)rat model was observed,and part of the treatment mechanism of Huoxue Jiedu Huayu Recipe on chronic kidney disease(CKD)was investigated.Methods Forty male wistar rats were randomly divided into 4 groups:the sham operation(Sham)group,the unilateral ureteral obstruction(UUO)group,the eplerenone treatment(EPL)group and the Huoxue Jiedu Huayu Recipe intervention(HJHR)group,10 rats in each group.The EPL group was given eplerenone and the HJHR group was given traditional Chinese medicine.The right kidney was collected after 6 months.Pathological changes were assessed by HE staining and Masson staining.The expressions of Cysteinyl aspartate specific proteinase1(Caspase-1),Interleukin-1β(IL-1β),Serum and glucocorticoid-induced kinase 1(SGK1)and nuclear factor κappa-B(NF-κB)were detected by immunohistochemistry.The DNA damage in renal VSMC was examined by TUNEL staining.Rat VSMCs were used for in vitro experiments.The cells were divided into 4 groups using the random number method:the control(CON)group,the aldosterone(ALD)group,the aldosterone plus eplerenone treatment(EPL)group and the aldosterone plus Huoxue Jiedu Huayu Recipe treatment(HJHR)group.The ALD group was given aldosterone stimulation for 24 h.The EPL group and the HJHR group were given eplerenone and rat serum containing 10%Huoxue Jiedu Huayu Recipe pretreatment before aldosterone stimulation,respectively.The apoptosis rate was detected by flow cytometry.Perforation of gasdermin D(GSDMD)and nuclear translocation of NR3C2(Encode MR)in cell membranes were examined by immunofluorescence.The protein expression of inflammation and pyroptosis-related signaling molecules were examined by western blotting.The mRNA expression of pyroptosis signaling pathway were detected by qRT-PCR.Results Compared with the Sham group,DNA damage of renal blood vessels and VSMCs was increased in UUO group,and expressions of Caspase-1,IL-1β,SGK1 and NF-κB were increased.Compared with the UUO group,the DNA damage of renal blood vessels and VSMCs in EPL and HJHR groups was reduced,and the expressions of Caspase-1,IL-1β,SGK1 and NF-κB were decreased.Compared with the CON group,the ALD group showed pyroptosis and DNA damage of rat VSMC.The expression of NR3C2,NLRP3,Caspase-1,GSDMD,IL-1β,SGK1 and NF-κB was up-regulated.Compared with the ALD group,the pyroptosis and DNA damage were alleviated,and the expression of NR3C2,NLRP3,Caspase-1,GSDMD,IL-1β,SGK1 and NF-κB was decreased in EPL and HJHR groups.Conclusion Huoxue Jiedu Huayu Recipe reduce the vascular damage of the contralateral kidneys of UUO by inhibiting the activation of the MR/NLRP3 inflammasome pathway and inhibiting the pyroptosis of VSMC.

OBJECTIVE To analyze the research status， hotspots， and trend of the clinical comprehensive evaluation of Chinese patent medicine in China. METHODS Based on CNKI， VIP and Wanfang database， clinical comprehensive evaluation of Chinese patent medicine was used as the subject of retrieval， and the retrieval time was from the inception to October 30th， 2022. CiteSpace 6.1.R3 and VOSviewer were used to conduct a visualization analysis of the relative literature of clinical comprehensive evaluation of Chinese patent medicine in terms of annual publication quantity， authors， institutions， keywords， etc. RESULTS & CONCLUSIONS A total of 1 460 pieces of literature related to the clinical comprehensive evaluation of Chinese patent medicine were included. The overall annual publication quantity showed a growth trend. There were 714 authors in the included literature. The institutions with a large publication quantity included the Chinese Academy of Traditional Chinese Medicine， Beijing University of Chinese Medicine， Tianjin University of Traditional Chinese Medicine， etc.， and there was little cooperation among the authors and institutions. High-frequency keywords included Chinese patent medicine， safety， adverse drug reactions， rational drug use， etc. Research hotspots focused on the safety and effectiveness evaluation of Chinese patent medicine. It may be a research trend in this field to strengthen the prescription review of Chinese patent medicine， and build a multi-dimensional and multi-criteriaclinical comprehensive evaluation system for the rational use of Chinese patent medicine.

Objective:To evaluate the therapeutic effect of hemopurification on acute chlorfenapyr poisoning according to the blood concentration of chlorfenapyr and to provide experience for clinical treatment.Methods:Two patients who presented to our Emergency Department following an ingestion of chlorfenapyr and then were treated with hemopurification in 2022 were included. The concentrations of chlorfenapyr and its highly toxic metabolite tralopyril were dynamically monitored, and the clinical data of the patients were collected.Results:Case 1 was given hemoperfusion for the first time 13 hours after ingestion. During l hour hemoperfusion, the tralopyril decreased by 28.82%. The concentration increased and exceeded the pre-perfusion level after 2 hours of hemoperfusion. After three times of hemoperfusion, the concentrations of chlorfenapyr and tralopyril were still higher than those before the first time, reaching 248 ng/mL and 1 307 ng/mL respectively. The concentration of chlorfenapyr showed a downward trend after 130 h, and the tralopyril in blood reached the peak 3 164 ng/mL at 130 h and decreased to 2 707 ng/mL at 178 h. In case 2, the blood chlorfenapyr and tralopyril concentration was 392 ng/mL and 7 598 ng/mL respectively 150 hours after ingestion. The blood chlorfenapyr concentration decreased by 37.75% respectively after first hemoperfusion, and the tralopyril concentration decreased by 38.02% respectively. During 85 hours of continuous veno-venous hemodiafiltration (CVVHDF), the concentration of tralopyril was maintained at 4 234~6 410 ng/mL. Case 1 was followed up to 12 days and lost follow-up. Case 2 died and the survival time was 247 hours.Conclusions:Hemoperfusion can scavenge tralopyril, but CVVHDF has poor scavenging ability for tralopyril. And the apparent volume of distribution (Vd) of chlorfenapyr and tralopyril are large. After ingestion, chlorfenapyr spreads to various tissues quickly, and it is easy to accumulate in the adipose tissue. The chlorfenapyr in the tissue slowly is released back to the blood and stays in the blood for a long time. The peak concentration of chlorfenapyr appeared earlier than that of tralopyril. Clinicians should pay attention to the early removal of toxins from the digestive tract.

Objective:To describe the current situation of gastric lavage operation and put forward measures for improvement by analyzing the clinical characteristics of 294 patients with gastric lavage in Poisoning Treatment Center of The First Affiliated Hospital of Nanjing Medical University.Methods:The clinical data of 294 patients with acute poisoning and gastric lavage from 2019 to 2021 were collected and analyzed retrospectively, and the related parameters (poison type, gastric lavage volume, poisoning to gastric lavage time, etc.) of each year were compared.Results:A total of 653 poisoning patients underwent gastric lavage from 2019 to 2021, with an average age of (44.2 ±20.1) years, and 134 (45.6%) were male. The main causes of gastric lavage were pesticide poisoning (52.72%) and drug poisoning (42.86%). The volume of gastric lavage was less than 10 L for 43.8% of patients and 10-20 L for 32.7% of patients. Patients with gastric lavage within 60 min after ingestion of poison accounted for 45.3%, followed by 25.8% within 61-120 min. The in-hospital mortality rate was 17.7%. The common complications of gastric lavage were: the incidence of gastrointestinal bleeding (55/121, 45.5%), the incidence of aspiration pneumonia (54/140, 38.6%), and the incidences of electrolyte disorder (21% of low potassium, 29% low calcium, and 10.0% low sodium). Compared with the groups in different years, the proportion of gastric lavage in poisoning was 58.85% vs. 46.60% vs. 32.41%, which decreased year by year, with statistical difference ( P <0.05). And there was no difference in the period from ingestion to gastric lavage and gastric lavage fluid volume. There was an increasing trend in poison types between diquat and other insecticides, but there was no statistical difference. Conclusions:From 2019 to 2021, the most common causes of acute gastric lavage were pesticide poisoning and drug poisoning, and the proportion of diquat and other pesticides showed an overall upward trend. A majority of the patients (71.1%) had gastric lavage within 2 h, and 76.5% of the patients had less than 20 L gastric lavage fluid. In the future, we will further control the amount of gastric lavage fluid and pay attention to the gastric lavage operation of new insecticide poisoning.