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 assess the validity and reliability of the Chinese version of the eleven-item Kutcher Adolescent Depression Scale (KADS-11)in Chinese adolescents,calculate its optimal cut-off value and the sensi-tivity and specificity,and explore the possibility of providing a useful tool to assess the severity of adolescent de-pressive symptoms.Methods:Totally 3180 students aged 11 -17 years were selected from schools in 6 provinces and Shanghai.All of them were asked to complete the KADS-11 and Children Depression Inventory (CDI). Students whose CDI scores were above 19 (including 19)were diagnosed with the DSM-IV criteria of depressive disorder,73 students from Shanghai sample were assessed with KADS-11 and CDI to analyze the test-retest reliabil-ity 1 month later.Results:Exploratory factor analysis showed that KADS-11 had 2 factors,and confirmatory factor analysis tested proved the 2-factor model fit better than the one-factor model.The KADS-11 total scores were posi-tively correlated with CDI total scores (r =0.74,P <0.01 ),and the KADS-11 scores were higher in depressive group than those in non-depressive group.The mean area under the curve (AUC)of KADS-11was 0.94,the mean area under the curve of each item ranged from 0.7 to 0.9.The optimal cut-off point of KADS-11 was total score≥9,sensitivity and specificity were 89% and 90% respectively.The Cronbach's alpha coefficient of the KADS-11 was 0.84,the spilt-half reliability coefficient was 0.71 (P <0.01),and the test-retest coefficient was 0.77 (P <0.01).Conclusion:The KADS-11 is appropriate for Chinese adolescents because of its good validity,reliability and diagnosis accuracy,it could be used to assess depressive symptoms for adolescents.

Objective To investigate the effect of urinary training on urination control in infants using nappies after birth.Methods Stratified sampling method was used to investigate the daytime urination control and the using of diapers by children in kindergartens of 6 prefecture-level cities in Henan Province,China.The survey was conducted among parents of healthy children.Results In a total of 12 250 questionnaires,11 697 had response,and 10 562 valid questionnaires were collected,and the effective recovery rate was 86.22％.After the birth of infants,the age of toilet training was divided into 6 groups (0-＜ 3 months old group,3-＜ 6 months old group,6-＜ 12 months old group,12-＜ 18 months old group,18-＜ 24 months old group and no toilet training group).Results showed that toilet training within 12 months leads to higher urinary control rate compared with toilet training after 12 months and no toilet training till the age of 2 years old,70.56％ (4 831/6 847 cases) in contrast with 59.02％ (1 545/2 618 cases) and 42.48％ (466/1 097 cases),and the differences were statistically significant (x2 =114.76,335.48,all P ＜ 0.000 1).Within 12 months,there was no statistical difference in the urinary control rate between subgroups at 2 years of age(all P ＞0.05).After 12 months,the urination control rate decreased with the start time delayed till 2 years of age,and the urinary control rate at the age of 2 years old n different groups was 59.97％ (1 314/2 191 cases) and 54.10％ (231/427 cases) respectively.The results of other ages(0.5,1.0,1.5 years old) were similar to those of 2 years old.There was no difference between the groups of different genders (all P ＞ 0.05).Conclusions The use of diapers in infants before the age of 1 year to start urinary training is conducive to the development of urinary control in infants.

Macrophages are important innate immune cells. Under inflammatory stimulation, macrophages rapidly respond and subsequently produce large amounts of cellular metabolites through metabolic reprogramming. Itaconate is an immunomodulatory derivative from the tricarboxylic acid cycle which has antioxidative and anti-inflammatory effects. In recent years, it has been reported that itaconate promotes the transition of macrophage phenotype from M1 to M2 and the underlying mechanism may include the activation of nuclear factor E2-related factor 2 (Nrf2) by alkylation of Kelch-like ECH-associated protein 1 (Keap1), inhibition of succinate dehydrogenase (SDH) and reactive oxygen species (ROS), blockade of the inhibitor ζ of nuclear factor-κB (IκBζ) translation and inhibition of aerobic glycolysis. In this review, we describe the metabolic pathways of itaconate, clarify the relationship between itaconate and the immune response, and summarize the latest researches about the roles of itaconate on regulating the inflammatory response in macrophages in order to provide the basis for the clinical use of itaconate and new strategies for the treatment of inflammatory diseases.

Objective:To compare the clinical efficacy of unilateral biportal endoscopy (UBE) and uniportal endoscopy (UE) for unilateral laminotomy for bilateral decompression (ULBD) in the treatment of lumbar spinal stenosis.Methods:Data of 82 patients with lumbar spinal stenosis treated by ULBD under UBE or UE from January 2020 to June 2021 in Dalian Central Hospital affiliated to Dalian Medical University and the First Hospital affiliated to Wenzhou Medical University were retrospectively analyzed, including 36 males and 46 females, aged 63.3±7.5 years (range, 47-81 years). According to the surgical procedure, they were divided into UBE group (42 cases), including 20 males and 22 females; aged 63.2±7.6 years (range, 47-81 years) and UE group (40 cases), including 16 males and 24 females; aged 63.5±7.5 years (range, 48-80 years). Operation time, hospital stay and surgical complications were compared between the two groups. Visual analogue scale (VAS) of low back and leg pain before surgery, 1 day, 7 d, 1 month and 6 months after surgery, and Oswestry disability index (ODI) before surgery, 1 month and 6 months after surgery were compared. Dural sac area before and after surgery, resection angle of ipsilateral facet joint, decompression rate of disc space and bone lateral recess were calculated.Results:All patients were operated successfully. In the UBE group, the operation time was 63.1±7.0 min, and the hospital stay was 3.9±0.9 d. The UE group was 61.2±6.2 min and 3.7±0.9 d, respectively ( t=1.31, P=0.195; t=1.24, P=0.217). The VAS of back and legs pain in UBE group decreased from 7.19±0.97 before operation to 3.43±0.63 points at postoperative 1 day, 1.71±0.60 at postoperative 7 d, 1.33±0.48 at postoperative 1 month and 1.36±0.48 points at postoperative 6 months ( F=352.29, P<0.001). The VAS score of the UE group decreased from 6.85±0.89 points before operation to 2.45±0.75 points at postoperative 1 day, 1.75±0.59 points at postoperative 7 d, 1.33±0.47 points at postoperative 1 month and 1.28±0.45 points at postoperative 6 months ( F=291.44, P<0.001). The VAS of low back and leg pain was higher in the UBE group than in the UE group at 1 day postoperatively ( t=6.41, P<0.001), and the difference was not statistically significant at 7 d postoperatively ( t=-0.27, P=0.786). The ODI of UBE group decreased from 66.62%±4.98% before operation to 21.81%±2.61% at postoperative 1 month and 11.62%±2.31% at postoperative 6 months ( F=1991.35, P<0.001). The ODI score of UE group decreased from 64.35%±5.16% before operation to 22.85%±3.26% at postoperative 1 month and 11.15%±2.86% at postoperative 6 months ( F=1931.18, P<0.001). The postoperative dural sac area of the UBE and UE groups was 135.1±10.0 mm 2 and 120.9±10.4 mm 2 ( t=6.30, P<0.001). The resection angle of ipsilateral facet joint was 69.3°±4.9° and 94.3°±4.1° in the two groups, respectively, with a statistically significant difference ( t=-25.00, P<0.001). The decompression rate of ipsilateral disk-flavum space was 39.0%±3.0% and 38.7%±3.3% in the two groups ( t=1.52, P=0.314). On the contralateral side was 41.6%±3.3% and 22.8%±3.2% ( t=26.32, P<0.001), respectively. The ipsilateral osseous side fossa decompression rate in the two groups were 70.0%±4.8% and 59.3%±3.9% ( t=15.64, P<0.001), the contralateral were 73.0%±3.4% and 48.4%±4.3% ( t=28.86, P<0.001). There was no significant difference in the decompression rate of ipsilateral disco-flavum space or bony lateral recess between the UBE group and the contralateral group ( t=-1.40, P=0.174; t=-1.72, P=0.096), while the decompression rate of discoflavum space and bony side recess on the ipsilateral side of UE group were higher than those on the contralateral side ( t=28.51, P<0.001; t=13.95, P<0.001). Conclusion:Both UE-ULBD and UBE-ULBD have good short-term clinical efficacy in patients with lumbar spinal stenosis. UB is better than UBE in early postoperative pain relief. However, UBE shows better imaging performance in decompression effect and better retention of facet joints.

Objective:To investigate the current situation of emergency medical service (EMS) system and its effect on treatment of the acute stage and short- and long-term prognosis in patients with acute myocardial infarction in Hebei province.Methods:Totally 2 961 patients with acute myocardial infarction who were admitted to major tertiary and some representative secondary hospitals in Hebei province from January 2016 to December 2016 were collected. According to the pattern of arriving hospital, all the patients were divided into the EMS group and self-transport group. The general conditions, time from onset to treatment, treatment methods, in-hospital mortality rate and 3-year mortality rate were compared between the two groups.Results:Of the included 2 961 patients, 33.13% of them were transported through EMS and 66.87% of them by private transport. Patients with a history of hypertension and ST-segment elevation myocardial infarction were more likely to choose EMS, and the difference was statistically significant ( P<0.05). Moreover, patients in the EMS group were more likely to go to tertiary hospitals for treatment (88.58% vs 85.76%, P=0.033). The time from onset to treatment of the EMS group was significantly shorter than that of the self-transport group (160 min vs 185 min, P<0.01), and the proportion of patients in the EMS group from onset-to-door time in <3 h and 3-6 h was higher than that of the self-transport group (55.76% vs 49.14%, 21.41% vs 19.09%, P<0.01). Compared with the self-transport group, the EMS group has a higher rate of reperfusion therapy (67.48% vs 61.67%, P=0.002). Patients in the EMS group had a higher in-hospital mortality rate in the acute stage (7.03% vs 4.44%, P=0.003), but its 3-year mortality rate was lower than that of the self-transport group (17.31% vs 20.77%, P<0.05). Conclusions:EMS can shorten symptom-onset-to-arrival time, increase the rate of reperfusion therapy and improve long-term prognosis of patients with acute myocardial infarction.

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.

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.