Homo- or heterodimeric compounds that affect dimeric protein function through interaction between monomeric moieties and protein subunits can serve as valuable sources of potent and selective drug candidates. Here, we screened an in-house dimeric natural product collection, and panepocyclinol A (PecA) emerged as a selective and potent STAT3 inhibitor with profound anti-tumor efficacy. Through cross-linking C712/C718 residues in separate STAT3 monomers with two distinct Michael receptors, PecA inhibits STAT3 DNA binding affinity and transcription activity. Molecular dynamics simulation reveals the key conformation changes of STAT3 dimers upon the di-covalent binding with PecA that abolishes its DNA interactions. Furthermore, PecA exhibits high efficacy against anaplastic large T cell lymphoma in vitro and in vivo, especially those with constitutively activated STAT3 or STAT3^Y640F. In summary, our study describes a distinct and effective di-covalent modification for the dimeric compound PecA to disrupt STAT3 function.

ObjectiveTo investigate the clinical features of patients with hepatolenticular degeneration （HLD） aged above 35 years. MethodsA retrospective analysis was performed for the clinical data of the patients with HLD， aged above 35 years， who attended West China School of Public Health， Sichuan University， from April 2018 to April 2023， and according to their clinical symptoms， they were divided into mixed type group with 13 patients， liver type group with 12 patients， and brain type group with 5 patients. Related data were collected， including general information （sex， clinical manifestation， age at confirmed diagnosis， time from initial symptoms to confirmed diagnosis， and family history）， laboratory examination （routine blood test， liver and renal function， serum copper， serum ceruloplasmin， urinary copper， and coagulation function）， and radiological examination. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups； the Fisher’s exact test was used for comparison of categorical data between groups. ResultsFor the 30 patients with HLD， the male/female ratio was 3∶1， and the mean age was 46.13±5.88 years； the patients with positive Kayser-Fleischer ring of the cornea accounted for 43.33%， and the patients with liver cirrhosis accounted for 66.67%. There were significant differences between the three groups in globulin， albumin/globulin ratio， alanine aminotransferase， prothrombin time， international normalized ratio， and activated partial thromboplastin time （F=5.893， 4.513， 4.424， 5.029， 5.248， and 4.942， all P<0.05）. ConclusionMost patients are male among the patients diagnosed with HLD after 35 years of age， with the main clinical types of mixed type and liver type， and such patients tend to have poor liver and coagulation functions. For unexplained liver function abnormalities and liver cirrhosis in this age group， the indicators such as serum ceruloplasmin and urinary copper should be screened as early as possible， and liver and kidney function and coagulation function should be monitored.

Objective To explore the protective effect and mechanism of Jinyinqingre oral liquid on acute lung injury in-duced by lipopolysaccharide(LPS)in mice.Methods C57BL/6J mice were randomly divided into six groups according to the random number table method:blank control group,model control group,Jinyinqingre oral liquid low-dose group,Jinyinqingre oral liquid medium-dose group,Jinyinqingre oral liquid high-dose group,and dexamethasone group.Except for the blank control group,the other groups were injected with lipopolysac charide(LPS)(5 mg·kg-1)into the trachea to establish the acute lung injury model of mice,and the Jinyinqingre oral liquid low,medium,and high groups were continuously administered the drug by gavage for three days.After 24 h,lung tissue and bronchoalveolar lavage fluid(BALF)were collected from the six groups for follow-up detection.The pathological injury of lung tissue in each group was observed by HE staining.The total number of cells in BALF was detected.The to-tal protein content of BALF was detected by the BCA method.The contents of inflammatory cytokines TNF-α,IL-1β,IL-6,and IgM in BALF were detected by ELISA.The expression of NF-κB and NLRP3 proteins in lung tissues was detected by immunohistochemistry and Western blotting.Results Compared with model control group,Jinyinqingre oral liquid alleviated the pathological injury of lung tissue(P＜0.05),decreased the total cell count,total protein content and IgM expression in BALF(P＜0.01),and the expres-sion of inflammatory cytokines TNF-α,IL-6 and IL-1β in BALF was dreased(P＜0.05),the protein expressions of NF-κB and NL-RP3 in lung tissues was dreased(P＜0.05).Conclusion Jinyinqingre oral liquid attenuated the pathological injury,inflammatory exudation,and expression of inflammatory cytokines in LPS-induced lung injury in mice,and its mechanism may be through the reg-ulation of NF-κB/NLRP3 signaling pathway,providing a theoretical basis for its clinical application.

Objective:To investigate the effect of methylene blue (MB) on motor dysfunction and its mechanism in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) mouse models.Methods:Forty healthy male C57BL/6 mice were randomly divided into 4 groups: control group, model group, low-dose treatment group and medium-dose treatment group ( n=10); PD mouse models were established by intraperitoneal injection of 25 mg/kg/d MPTP for a consecutive 7 d; low-dose treatment group and medium-dose treatment group were pretreated intraperitoneally with MB 2 mg/kg/d or MB 10 mg/kg/d for a consecutive 3 d, respectively; and then, MPTP 25 mg/kg/d+MB 2 mg/kg/d or MPTP 25 mg/kg/d+MB 10 mg/kg/d were injected intraperitoneally into the low-dose treatment group or medium-dose treatment group for a consecutive 7 d (MPTP and MB were given at 12 h of interval). Eight d after modeling, open field experiment, pole climbing experiment and rod rotating experiment were carried out to evaluate the spontaneous movement, coordination, endurance and motor ability. And then, the mice were sacrificed; immunofluorescent staining was used to observe tyrosine hydroxylase (TH) expression in the substantia nigra; Western blotting was used to detect the expressions of TH, α-synuclein, nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-Caspase-1 and Gasdermin D (GSDMD) in the striatum and substantia nigra of mice. Contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-18 in the substantia nigra and striatum of mice were detected by ELISA. Results:Compared with the control group, the model group had shortened residence time in rod rotating, prolonged descent time in rod climbing, reduced total movement distance in open field, decreased number of TH-positive cells in the substania nigra, decreased TH protein levels in the substania nigra and striatum, and increased NLRP3, ASC, cleaved-Caspase-1, GSDMD and GSDMD-N protein levels in the substania nigra and striatum, and increased TNF-α, IL-1β and IL-18 contents in the substania nigra and striatum, with significant differences ( P<0.05). Compared with the model group, low-dose treatment group and medium-dose treatment group had prolonged residence time in rod rotating, shortened descent time in rod climbing, increased total movement distance in open field, increased number of TH-positive cells in the substania nigra, and increased TH protein levels in the substania nigra and striatum, decreased NLRP3, ASC, and cleaved-Caspase-1 levels in the substania nigra and striatum, and decreased TNF-α, IL-1β and IL-18 contents in the substania nigra and striatum, with significant differences ( P<0.05). No statistical differences in the above indexes were noted between the low-dose treatment group and medium-dose treatment group ( P>0.05). Conclusion:Low-/medium-dose MB can ameliorate motor dysfunction in PD mouse models, whose mechanism may be related to downregulate NLRP3 inflammasome and inhibit neuroinflammatory response to reduce dopaminergic neuron pyroptosis.

BACKGROUND: Sweat secreted by eccrine sweat glands is transported to the skin surface through the lumen. The eccrine sweat gland develops from the initial solid bud to the final gland structure with a lumen, but how the lumen is formed and the mechanism of lumen formation have not yet been fully elucidated. This study aimed to investigate the mechanism of lumen formation of eccrine gland organoids (EGOs). METHODS: Human eccrine sweat glands were isolated from the skin for tissue culture, and the primary cultured cells were collected and cultured in Matrigel for 14 days in vitro. EGOs at different development days were collected for hematoxylin and eosin (H&E) staining to observe morphological changes and for immunofluorescence staining of proliferation marker Ki67, cellular motility marker filamentous actin (F-actin), and autophagy marker LC3B. Western blotting was used to detect the expression of Ki67, F-actin, and LC3B. Moreover, apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assay kit, and the expression of poly (ADP-ribose) polymerase and Caspase-3 was detected by Western blot. In addition, 3-methyladenine (3MA) was used as an autophagy inhibitor to detect whether the formation of sweat glands can be effectively inhibited. RESULTS: The results showed that a single gland cell proliferated rapidly and formed EGOs on day 4. The earliest lumen formation was observed on day 6. From day 8 to day 14, the rate of lumen formation in EGOs increased significantly. The immunofluorescence and Western blot analyses showed that the expression of Ki67 gradually decreased with the increase in days, while the F-actin expression level did not change. Notably, the expression of autophagy marker LC3B was detected in the interior cells of EGOs as the apoptosis signal of EGOs was negative. Compared with the control group, the autophagy inhibitor 3MA can effectively limit the formation rate of the lumen and reduce the inner diameter of EGOs. CONCLUSION Using our model of eccrine gland 3D-reconstruction in Matrigel, we determined that autophagy rather than apoptosis plays a role in the lumen formation of EGOs.
Apoptosis ; Autophagy ; Eccrine Glands ; Epithelial Cells ; Humans ; Organoids

Objective:To compare preoperative portal vein embolization (PVE) using tris-acryl gelatin microspheres (TAGM) versus coils.Methods:From March 2016 to June 2018, 21 consecutive patients with a future liver remnant (FLR) ratio of less than 45% before planned major hepatectomy for malignant or benign liver diseases were enrolled from the First Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital into this study. The patients were divided to receive portal vein embolization (PVE) using TAGM distally and coils proximally (the TC group) and PVE using multiple coils (the CC group). Post-PVE complications, liver function, routine blood tests; FLR hyperplasia, types of liver resection, operation time, intraoperative blood loss, and postoperative complications were compared between the two groups.Results:Eight patients were included in the TC group. There were 4 males and 4 females, with a mean age of (55.3±7.7) years. Of 13 patients included into the CC group, there were 11 males and 2 females, with a mean age of (52.6±11.3) years. There were no significant differences in sex, age, types of hepatic diseases, volume of FLR, ratio of FLR, ratio of standard FLR, types of surgery, operation duration, blood loss, major complications, and liver failure rates between the two groups. All patients in the two groups had successful PVE. The TC group developed effective growth of volume of FLR with one patient who failed to undergo surgery because of tumor progression. In the CC group, four patients failed to undergo liver resection: one patient developed thrombosis of the left branch and main trunk of portal vein; tumor progression occurred in one patient and two patients had insufficient FLR growth. Compared with the CC group, the TC group had a significantly higher volume of FLR hyperplasia [(9.0±2.8) % vs. (5.2±3.8) %, P<0.05], and a faster but insignificant increase in proliferation rate [(11.4±7.1) ml/d vs. (6.9±5.2) ml/d, P>0.05], a greater but insignificant increase in percentage of proliferation [(33.6±20.1) % vs. (20.9±15.1) %, P>0.05]. Conclusions:This study showed that PVE with TAGM plus coils is safe and effective. It induced a better degree of hypertrophy of FLR compared to PVE using multiple coils.

Objective To investigate the intervention of family doctor management system on the loneliness in people who lost their only child.Methods Sixty two residents who lost their only child in Shanghai Xujiahui district were enrolled in the study from December 2015 to June 2016.The participants were randomly divided into study group and control group with 31 cases in each group;the participants in study group were signed by family doctors and the psychological intervention was provided,those in control group had no intervention,the participants were followed up for 6 months,the psychological status was evaluated with UCLA loneliness scale in two groups.Results There were no significant differences in UCLA scores (63.26 ±7.17 vs.60.03 ±7.59,t =1.709,P ＞0.05) and in the severity of loneliness (mild 0/30 vs.1/31,moderate 9/30 vs.11/31,severe 21/30 vs.19/31,x2 =1.284,P ＞ 0.05) between study and control groups before the intervention.There were significant differences in UCLA scores (63.26 ±7.17 vs.40.13 ± 5.61,t =15.631,P ＜ 0.05) and in the severity of loneliness (mild 0/30 vs.17/30,moderate 9/30 vs.13/30,severe 21/30 vs.0/30,x2 =38.727,P ＜ 0.05) before and after 6 months of intervention in study group;while there were no significant changes in UCLA score (60.03 ± 7.59 vs.59.64 ± 6.90,t =0.650,P ＞ 0.05) and in the severity of loneliness (mild 1/31 vs.0/31,moderate 11/31 vs.14/31,severe 19/31 vs.17/30,x2 =1.471,P ＞ 0.05) in control group after 6-month follow-up.The UCLA scores (40.13 ±5.61 vs.59.64 ±6.90,t =12.086,P＜0.05) and the severity of loneliness (mild 17/30 vs.0/31,moderate 13/30 vs.14/31,severe 0/30 vs.17/31,x2 =34.030,P ＜ 0.05) in study group were significantly lower than those in control group after intervention.Conclusion The family doctor management system can improve the loneliness and mental health status of people who lost the only child,to reduce the risk of mental illness and the family and social burden.

Objective To evaluate the therapeutic effect of antibiotics combined with recombinant tuberculosis vaccine AEC/BC02 on Mycobacterium tuberculosis infection in guinea pigs. Methods Two weeks after guinea pigs were challenged subcutaneously with a high dose of Mycobacterium tuberculosis,the guinea pigs with the positive skin test responses to the recombinant ESAT6-CFP10 allergen were randomly di-vided into four groups:normal saline (NS),AEC/BC02,antibiotics and antibiotics+AEC/BC02. In antibiotics+AEC/BC02 group,guinea pigs firstly received isoniazid ( INH) and rifapentine ( RFT) treatment once a week for a total of three times,and then were immunized with a single dose of AEC/BC02 vaccine six times at 10-day intervals. Guinea pigs in AEC/BC02 and antibiotics groups were respectively vaccinated with AEC/BC02 vaccine and given INH and RFT treatment at the same dose and frequency as given to antibiotics+ AEC/BC02 group. Thirteen weeks after challenge,all guinea pigs were sacrificed for necropsy. Results The gross pathological scores of NS,AEC/BC02,antibiotics and antibiotics+AEC/BC02 groups were 83±8,77± 22,45±28 and 19±14,respectively. Antibiotics+AEC/BC02 group had a significantly lower gross pathological score than antibiotics,AEC/BC02 and NS groups (P<0. 05,P<0. 01,P<0. 01,respectively). Moreover,the gross pathological scores of antibiotics and AEC/BC02 groups were significantly decreased as compared with that of NS group (both P<0. 01). However,there was no significant difference between AEC/BC02 and NS groups. The spleen bacterial load of antibiotics+AEC/BC02 group was (2. 50±1. 26) lg CFU,which was sig-nificantly lower than those of NS [(4. 92+0. 52) lg CFU],AEC/BC02 [(4. 78+0. 84) lg CFU] and antibi-otics [(4. 39+0. 50) lg CFU] groups (P<0. 01). Compared with NS group,antibiotic and AEC/BC02 groups showed no significant difference in spleen bacterial load. Histopathological changes indicated different levels of granulomatous lesions appeared in lung tissues of all groups and the most severe change was ob-served in AEC/BC02 group,followed by that in NS,antibiotics and antibiotics+AEC/BC02 groups. Conclu-sion INH and RFT treatment in combination with AEC/BC02 vaccine in the treatment of guinea pigs with Mycobacterium tuberculosis infection was superior to either treatment alone as it significantly alleviated organ lesions and lowered the bacterial loads in spleen and lung.

Objective To evaluate the feasibility and efficacy of preoperative sequential transcatheter arterial chemoembolization (TACE) followed by selective portal venous embolization (PVE) in patients with marginally resectable hepatocellular carcinoma (HCC).The aim was to find out whether this combined procedure helped to increase the rate of extended radical liver resection.Methods From March 2009 to November 2016,29 patients with HCC which were marginally resectable underwent preoperative TACE combined with PVE were included into this study.All these patients were subsequently assessed to undergo radical hepatectomy.The complications,laboratory results,volume changes of each liver lobe and patient survival were analyzed.Results TACE combined with PVE was successful in all the 29 patients.There were no major complications.After the procedure,the volumes of the tumor and the part of the liver to be resected decreased to certain degree.The remnant liver volume (RLV) increased remarkably.The RLV were (395.4 ±58.7) cm3 and (599.2 ±75.2) cm3 before and after the procedure,respectively.The difference was significant (P ＜ 0.05).19 patients underwent radical hemihepatectomy or trisectionectomy,with a resection rate of 65.5％ (19/29).There were sufficient surgical margins in all the resected tumors.After operation,the 1-,3-,and 5-year survival rates were 58.8％,35.5％ and 17.6％,respectively.Conclusion For HCC patients who had marginally resectable HCC,preoperative TACE combined with PVE efficiently controlled the growth of the tumors,decreased the volume of the liver lobe with tumor,increased the RLV,and made it possible for a planned two-stage radical hepatectomy with sufficient surgical margin and better survival in a significant proportion of patients.

Genetic Therapy ; Hemophilia A ; therapy ; Humans