As activated hepatic stellate cells (aHSCs) play a central role in fibrogenesis, they have become key target cells for anti-fibrotic treatment. Nevertheless, the therapeutic efficiency is constrained by the exosomes they secrete, which are linked to energy metabolism and continuously stimulate the activation of neighboring quiescent hepatic stellate cells (qHSCs). Herein, an intercellular communication interference strategy is designed utilizing paeoniflorin (PF) loaded and hyaluronic acid (HA) coated copper-doped ZIF-8 (PF@HA-Cu/ZIF-8, PF@HCZ) to reduce energy-related exosome secretion from aHSCs, thus preserving neighboring qHSCs in a quiescent state. Simultaneously, the released copper and zinc ions disrupt key enzymes involved in glycolysis to reduce bioenergy synthesis in aHSCs, thereby promoting the reversion of aHSCs to a quiescent state and further decreasing exosome secretion. Therefore, PF@HCZ can effectively sustain both aHSCs and qHSCs in a metabolically dormant state to ultimately alleviate liver fibrosis. The study provides an enlightening strategy for interrupting exosome-mediated intercellular communication and remodeling the energy metabolic status of HSCs with boosted antifibrogenic activity.

ObjectiveTo observe the clinical effectiveness and safety of Xuandi Ziyin Mixture （玄地滋阴合剂） for central precocious puberty （CPP） in girls with syndrome of yin deficiency and fire exuberance， and to analyse the effect of body mass index （BMI） on the effectiveness. MethodsA total of 236 girls with CPP of yin deficiency and fire exuberance syndrome were included， and all of them were given Xuandi Ziyin Mixture， 30 ml each time， twice a day， for a total treatment period of 6 months. Before and after treatment， children's weight， height and bone age were measured， BMI and BMI Z-score （BMI Z） and the difference between bone age and actual age were calculated； ultrasound was used to detect uterine and ovarian sizes， and to calculate uterine volume （V_uterus）， bilateral ovarian volume （V_{left ovary}， V_{right ovary}）， and bilateral maximal follicle diameters （r_{left follicle} and r_{right follicle}）； and serum sex hormones were measured， including follicle-stimulating hormone （FSH）， luteinising hormone （LH）， prolactin （PRL）， estradiol （E₂）， and testosterone （T）， and were scored for traditional Chinese medicine （TCM） syndrome. Multiple linear regression was used to analyse the influence factors of the difference between bone age and actual age， and changes in uterine volume. The children were divided into the normal weight group and the overweight/obesity group according to baseline BMI， and the bone age， the difference between bone age and actual age， V_uterus and BMI Z scores before and after treatment were compared between the two groups. ResultsFinally， 199 children entered the statistical analysis. Compared with pre-treatment， the bone age， BMI and BMI Z scores of the children increased after treatment， and the difference between bone age and actual age， TCM syndrome scores， V_uterus， V_{left ovary}， V_{right ovary}， r_{left follicle} and r_{right follicle} decreased； and the levels of serum FSH， LH， E₂， and T significantly decreased （P＜0.05 or P＜0.01）. The difference between bone age and actual age was negatively correlated with LH and V_uterus （P＜0.05）， and changes in uterine volume were positively correlated with LH （P＜0.01）. Comparing between the groups before and after treatment， the bone age， difference between bone age and actual age， and BMI Z scores of children in the normal weight group （100 cases） were significantly smaller than those in the overweight/obesity group （99 cases） （P＜0.01）. Compared with pre-treatment， the bone age of the children in both groups increased， but the difference between bone age and actual age and V_uterus were significantly smaller （P＜0.01）. Further comparison of Δ bone age and actual age difference and ΔV_uterus （Δ = post-treatment value - pre-treatment value） between the two groups showed no statistically significant difference （P＞0.05）. ConclusionsXuandi Ziyin Mixture can reduce bone age， sex hormone level， ovary and uterus size， and improve clinical symptoms in girls with yin deficiency and fire exuberance syndrome of CPP. It is also effective in overweight/obese children， and the inhibitory effect on bone age in girls with CPP is not affected by BMI， with good safety.

The Ly-6 and uPAR (LU) domain-containing proteins represent a large family of cell-surface markers. In particular, mouse Ly-6A/Sca-1 is a widely used marker for various stem cells; however, its human ortholog is missing. In this study, based on a systematic survey and comparative genomic study of mouse and human LU domain-containing proteins, we identified a previously unannotated human gene encoding the candidate ortholog of mouse Ly-6A/Sca-1. This gene, hereby named LY6A, reversely overlaps with a lncRNA gene in the majority of exonic sequences. We found that LY6A is aberrantly expressed in pituitary tumors, but not in normal pituitary tissues, and may contribute to tumorigenesis. Similar to mouse Ly-6A/Sca-1, human LY6A is also upregulated by interferon, suggesting a conserved transcriptional regulatory mechanism between humans and mice. We cloned the full-length LY6A cDNA, whose encoded protein sequence, domain architecture, and exon-intron structures are all well conserved with mouse Ly-6A/Sca-1. Ectopic expression of the LY6A protein in cells demonstrates that it acts the same as mouse Ly-6A/Sca-1 in their processing and glycosylphosphatidylinositol anchoring to the cell membrane. Collectively, these studies unveil a novel human gene encoding a candidate biomarker and provide an interesting model gene for studying gene regulatory and evolutionary mechanisms.
Humans ; Membrane Proteins/genetics* ; Pituitary Neoplasms/genetics* ; Biomarkers

Anti-CD19 chimeric antigen receptor (CAR)-T cell therapy has achieved 40%-50% long-term complete response in relapsed or refractory diffuse large B-cell lymphoma (DLBCL) patients. However, the underlying mechanism of alterations in the tumor microenvironments resulting in CAR-T cell therapy failure needs further investigation. A multi-center phase I/II trial of anti-CD19 CD28z CAR-T (FKC876, ChiCTR1800019661) was conducted. Among 22 evaluable DLBCL patients, seven achieved complete remission, 10 experienced partial remissions, while four had stable disease by day 29. Single-cell RNA sequencing results were obtained from core needle biopsy tumor samples collected from long-term complete remission and early-progressed patients, and compared at different stages of treatment. M2-subtype macrophages were significantly involved in both in vivo and in vitro anti-tumor functions of CAR-T cells, leading to CAR-T cell therapy failure and disease progression in DLBCL. Immunosuppressive tumor microenvironments persisted before CAR-T cell therapy, during both cell expansion and disease progression, which could not be altered by infiltrating CAR-T cells. Aberrant metabolism profile of M2-subtype macrophages and those of dysfunctional T cells also contributed to the immunosuppressive tumor microenvironments. Thus, our findings provided a clinical rationale for targeting tumor microenvironments and reprogramming immune cell metabolism as effective therapeutic strategies to prevent lymphoma relapse in future designs of CAR-T cell therapy.

OBJECTIVE To investigate the effects of omeprazole on pharmacokinetic parameters of imatinib in rats. METHODS According to body weight， the rats were divided into imatinib+low-dose， medium-dose， and high-dose omeprazole groups， imatinib group， with 6 rats in each group. They were given omeprazole suspension at the doses of 1.8， 3.6 and 7.2 g/kg， or 0.5% sodium carboxymethyl cellulose solution intragastrically respectively； one hour later， imatinib suspension was administered by oral gavage at a the dose of 10 mg/kg. Blood sample （100 μL） was taken from the orbit before and 0.5， 1， 2， 2.5， 3， 4， 5， 6， 8， 12， 24 and 36 hours after intragastric administration of imatinib. Using imatinib-d3 as internal standard， the plasma concentrations of imatinib and its metabolite N-desmethyl imatinib in rat were determined by high performance liquid chromatography-tandem mass spectrometry. The pharmacokinetic parameters were calculated by DAS 2.0 software and compared. RESULTS Compared with imatinib group， AUC0－∞ and AUMC0－∞ of imatinib in rat plasma of imatinib+medium-dose omeprazole group， cmax， t1/2， AUC0－∞ and AUMC0－∞ of imatinib in rat plasma of imatinib+high-dose omeprazole group were all increased or prolonged significantly （P＜0.05）. Compared with imatinib group， AUC0－∞ and AUMC0－∞ of N-desmethyl imatinib in rat plasma of imatinib+medium-dose omeprazole group， and cmax and AUC0→∞ of N-desmethyl imatinib in rat plasma of imatinib+high-dose omeprazole group were decreased significantly （P＜0.05）. CONCLUSIONS Omeprazole may increase the plasma concentration of imatinib in rats and reduce the plasma concentration of N-desmethyl imatinib in rats， which may be associated with inhibiting the metabolism of imatinib.

Objective:To investigate the development of intrapulmonary vascular volume (IPVV) in normal preschool children based on quantitative measurement on chest CT.Methods:The CT data of 407 normal preschool children (236 males and 171 females, aged 1-72 months, with a median of 36 months) who underwent chest CT examination from January 2014 to May 2017 in the "Digital Lung" imaging database were retrospectively collected. The pulmonary vessels were segmented by the "Digital Lung" automatic detection tool, and the IPVV of the whole lung, the right lung, the left lung and each lobe were obtained, and the IPVV upper/lower and IPVV left/right were calculated. According to the age, the subjects were divided into infant period (0-12 months), early childhood period (13-36 months) and preschool period (37-72 months), with 30 cases (17 males and 13 females), 175 cases (95 males and 80 females) and 202 cases (124 males and 78 females) respectively. Spearman correlation analysis was used to evaluate the correlation between IPVV and month age. One-way analysis of variance or Kruskal-Wallis H test was used to compare the differences of IPVV, IPVV upper/lower and IPVV left/right between different months of age. Independent sample t test or Mann-Whitney U test was used to compare the differences of IPVV between different genders, and the normal reference range of IPVV in normal preschool children of different months of age was established. Results:The IPVV of the whole lung, right lung, left lung and each lung lobe were positively correlated with age, the correlation coefficient was 0. 638-0.820 in males and 0. 683-0.791 in females (all P<0.001). There was no significant difference in IPVV of whole lung, right lung, left lung and each lobe between male and female from 0 to 12 months (all P>0.05), but there was significant difference in IPVV of whole lung, right lung, left lung and each lobe between male and female from 13 to 36 months (all P<0.05). There were significant differences in IPVV of the whole lung, right lung, left lung and upper lobe of both lungs between boys and girls from 37 to 72 months (all P<0.05). IPVV upper/lower in the right lung (χ 2=14.00, 12.87, P=0.001, 0.002) and IPVV upper/lower in the left lung (χ 2=6.65, 22.84, P=0.036,<0.001) were significantly different in both boys and girls among 3 months of age. And with the increase of age, it showed a decreasing trend. There was no significant difference in IPVV upper/lower between boys and girls at the same age (all P>0.05).There was no significant difference in IPVV left/right among different months and between different sexes (all P>0.05). Finally, the normal reference value range of IPVV of different genders in infancy, early childhood and preschool age was calculated. Conclusions:The increase of pulmonary vessels in normal preschool children is positively correlated with age. There is no significant difference in IPVV between boys and girls in infant period, but IPVV in boys is larger than that in girls in early childhood period and preschool period. IPVV in the lower lung increased faster than that in the upper lung, but there was no significant difference between the left and right lungs.

Objective:To investigate the characteristics of short-term prognostic factors in very elderly patients with acute coronary syndrome (ACS).Methods:A total of 2 912 ACS patients admitted to Shengjing Hospital of China Medical University from January 1, 2010 to October 31, 2014 and treated with percutaneous coronary intervention (PCI) were enrolled and divided into two groups according to age: very elderly group (≥75 years, 480 cases) and control group (< 75 years, 2 432 cases). The clinical data and coronary artery lesions of the included patients were detected. Major cardiovascular adverse events (MACE) occurred within 30 d after discharge were followed up and recorded. The short-term prognostic factors in very elderly patients with ACS were analyzed by Logistic regression.Results:Compared with control group, the percentage of hypertension, global registry of acute coronary events (GRACE) score, high density lipoprotein cholesterol, N-terminal pro-brain natriuretic peptide, the left main lesion ratio and Gensini score in very elderly group were higher, while the percentage of men, number of smoking, hyperlipidemia proportion, red blood cell count, white blood cell count, blood platelet count, albumin, and long term oral administration of aspirin, clopidogrel, statins, angiotensin receptor inhibitor after discharge were lower, and the differences were statistically significant ( P<0.01 or<0.05). During the follow-up period, the all-cause mortality in very elderly group was higher than that in control group: 2.5%(12/480) vs. 0.9% (21/2 432), and the difference was statistically significant ( P<0.01). Multivariate analysis showed that oral angiotensin converting enzyme inhibitor was a protective factor for elderly ACS patients after discharge ( OR = 0.046, 95% CI 0.006 to 0.383, P = 0.004). The receiver operating characteristic curve analysis showed that Gensini score ≥ 87.75 scores was a threshold value for all-cause mortality. The all-cause mortality ratio in high Gensini score (≥ 87.75 scores) group was higher than that in low Gensini score (<87.75 scores) group: 6.6% (9/137) vs. 0.9% (3/343), and the difference was statistically significant ( P<0.01). Conclusions:Very elderly patients with ACS have their own characteristics from both clinical history and prognostic factors. Patients with Gensini score of ≥ 87.75 scores should be closely observed, and drug treatment during hospitalization should be intensified if necessary. Follow-up should be strengthened for such patients, and oral drug treatment should be continued after discharge.

Objective:To compare the efficacy of bilayer artificial dermis graft plus single layer dermal template and bilayer artificial dermis graft only in repairing lower extremity wounds with large area of exposed bone.Methods:A retrospective case-control study was conducted to analyze the clinical data of 34 patients with 37 wounds of the lower extremity involving large area of exposed bone admitted to Beijing Jishuitan Hospital from November 2009 to November 2020. There were 27 males and 7 females, aged 9-67 years [35.5(29, 45)years]. The exposed bone in the lower leg, ankle and foot was greater than 10 cm 2 in size (the shortest distance from edge to edge of bony exposure was more than 2 cm). At the first stage, the wounds were grafted with bilayer type artificial dermis only for 21 wounds of 20 patients in Group A, and grafted with bilayer type artificial dermis plus single layer dermal template for 16 wounds of 14 patients in Group B. At the second stage, the auto-skin graft was performed in the two groups. The wound healing rate was observed in all patients, and was compared between the two groups at 2 weeks and 2 months after the second stage operation. At the same time, the interval between first stage and second stage surgery was measured. The Vancouver Scar Scale (VSS) was used to evaluate the scar in the skin grafting area in the two groups at 5-6 months after the second stage operation. Results:All patients were followed up for 1 to 24 months [5(2, 7.5)months]. The total excellent and good wound healing rate in all patients was 81%(30/37) at 2 weeks and 97%(36/37) at 2 months. There was no significant difference between the Group A and Group B in the excellent and good wound healing rate at 2 weeks [(76%(16/21) vs. 88%(14/16)] and at 2 months [95%(20/21) vs. 100%(16/16)] ( P>0.05). In Group A, the bilayer artificial dermis was grafted into 4 wounds again to complete exposed bone coverage. However, all wounds in Group B were covered initially without re-grafting. The interval between the two-stage operation was 20(16, 21)days in Group A after the 4 patients who underwent secondary artificial dermal transplantation were excluded, showing no significant difference from 21(21, 23)days in Group B ( P>0.05). At 5-6 months after the second stage operation, the VSS score in Group B [(8.0±1.2)points] was significant less than that in Group A [(9.2±1.1)days] ( P<0.05). In the sub-index of VSS, the score of color and softness of scar in Group B [(2.0±0.6)points, (1.6±0.5)points] were significantly improved compared to those in Group A [(2.5±0.5)points, (2.2±0.7)points] ( P<0.05). Conclusions:The artificial dermis grafting is effective in treatment of lower extremity wounds with large area of exposed bone. However, the bilayer artificial dermis graft plus single layer dermal template can avoid artificial dermal re-graft in repair of large area of exposed bone, and the interval between two-stage operation is not significantly prolonged. Moerover, the color and texture of scar after skin grafting and wound repair efficiency and quality are improved.

BACKGROUND: Hepatectomy for hepatocellular carcinoma (HCC) beyond the Milan criteria is shown to be beneficial. However, a high rate of post-operative HCC recurrence hinders the long-term survival of the patients. This study aimed to investigate and compare the impacts of tenofovir (TDF) and entecavir (ETV) on the recurrence of hepatitis B viral (HBV)-related HCC beyond the Milan criteria. METHODS: Data pertaining to 1532 patients who underwent hepatectomy and received antiviral therapy between January 2014 and January 2019 were collected from five centers. Recurrence-free survival (RFS) analysis was performed using the Kaplan-Meier method. Cox proportional hazards regression analysis was performed to determine prognostic factors for HCC recurrence. RESULTS: The analysis incorporates 595 HBV-related HCC patients. The overall 5-year RFS was 21.3%. Among them, 533 and 62 patients received ETV and TDF treatment, respectively. The 1-, 3-, and 5-year RFS rates were 46.3%, 27.4%, and 19.6%, respectively, in the ETV group compared with 65.1%, 41.8%, and 37.2%, respectively, in the TDF group (P < 0.001). Multivariate analysis showed that TDF treatment (hazard ratio [HR]: 0.604, P = 0.005), cirrhosis (HR: 1.557, P = 0.004), tumor size (HR: 1.037, P = 0.008), microvascular invasion (MVI) (HR: 1.403, P = 0.002), portal vein tumor thrombus (PVTT) (HR: 1.358, P = 0.012), capsular invasion (HR: 1.228, P = 0.040), and creatinine levels (CREA) (HR: 0.993, P = 0.031) were statistically significant prognostic factors associated with RFS. CONCLUSIONS Patients with HCC beyond the Milan criteria exhibited a high rate of HCC recurrence after hepatectomy. Compared to the ETV therapy, TDF administration significantly lowered the risk of HCC recurrence.
Antiviral Agents/therapeutic use* ; Carcinoma, Hepatocellular/surgery* ; Guanine/analogs & derivatives* ; Hepatectomy ; Hepatitis B virus ; Hepatitis B, Chronic/drug therapy* ; Humans ; Liver Neoplasms/surgery* ; Retrospective Studies ; Tenofovir/therapeutic use*

OBJECTIVE：To establish a method for the determination of pyrrotinib concentration in plasma ，and apply it in clinic. METHODS ：After precipitated with methanol ，the plasma sample was determined by LC-MS/MS using imatinib as internal standard. The determination was performed on Ultimate AQ-C 18 column with mobile phase consisted of methanol （containing 0.1％ formic acid ）and water （containing 0.1％ formic acid ）（gradient elution ）at the flow rate of 0.4 mL/min. The column temperature was 40 ℃，and the sample size was 5 µL. The ion source was electrospray ionization source ，and the positive ion scanning was carried out in multiple reaction mode. The ion pairs for quantitative analysis were m/z 583.4→138.3（pyrrotinib）and m/z 494.5→ 393.4（internal standard ），respectively. Thirty breast cancer patients taking pyrrotinib were collected from the Affiliated Hospital of Qingdao University during Jun.-Nov. 2020 to determine their steady-state trough concentrations of pyrrotinib after a week of treatment. RESULTS ：The linear range of pyrrotinib were 5-300 ng/mL（r＝0.999 3）. The lower limit of quantification was 5 ng/mL. RSDs of intra-day and inter-day were not higher than 9.30％，and relative errors （REs）ranged －6.70％-5.04％. REs of stability tests were in the range of －1.92％-5.42％. The extraction method ，matrix effect and residual effect did not affect the quantitative analysis of the substance to be tested. The steady-state trough concentrations of pyrrotinib were 32.6-82.8 ng/mL，with an average plasma concentration of 53.8 ng/mL；there was about 2.54 fold individual difference. CONCLUSIONS ：Established LC-MS/MS method is simple ，sensitive and accurate ，and can be used for the plasma concentration monitoring of pyrrotinib in breast cancer patient.