In this study, we constructed a series of recombinant Fc variants of immunoglobulin G4 (IgG4), screened the fragment crystallizable (Fc) variants with significantly prolonged serum half-lives, and analyzed the relationship between mutation site and half-life, aiming to provide a theoretical basis for the development of IgG4 antibodies and Fc fusion protein-based drugs. Nine gene sites were selected for mutation, and different mutation sites were combined. The variant expression plasmids pET24b-Fc were constructed by molecular cloning and point mutation. The plasmids were transformed into Escherichia coli BL21(DE3) for the expression of different recombinant proteins of Fc. Fc2 and Fc3 variants had slightly lower recombinant protein yields, and the expression of other variants was not affected. The toxicity of different Fc variants was determined by cell counting kit-8 (CCK-8) and calcein acetoxymethyl ester/ propidium iodide (calcein AM/PI) in vitro and enzyme-linked immuno sorbent assay (ELISA) in vivo. The results showed that the recombinant Fc variants had good biocompatibility and safety. Finally, the Fc variants were labeled with fluorescent markers, and the effects of different mutations on their serum half-lives were investigated by in vivo experiments. The Fc5 variant with prolonged serum half-life was successfully screened out, which provided a theoretical and practical basis for the optimal design of IgG4 subtype antibody and Fc fusion protein drugs.
Immunoglobulin G/blood* ; Immunoglobulin Fc Fragments/biosynthesis* ; Half-Life ; Animals ; Escherichia coli/metabolism* ; Humans ; Recombinant Fusion Proteins/biosynthesis* ; Recombinant Proteins/biosynthesis* ; Mice

Objective:To investigate the therapeutic value of visual endoscopic retrograde appendicitis therapy (vERAT) in pediatric patients with acute suppurative appendicitis (ASA).Methods:This was a retrospective cohort study. A total of 55 ASA patients who underwent vERAT at the Pediatric Department of the Tangdu Hospital of Air Force Medical University between November 2023 and January 2025 were selected and divided into groups based on the presence or absence of fecaliths: fecalith group and non-fecalith group. The baseline characteristics, initial treatment success rates, treatment costs, hospital stay duration, procedure time, and recurrence rates between two groups were compared. Mann-Whitney U test and χ2 test were used to evaluate group differences. Results:A total of 55 ASA patients were enrolled, including 38 males and 17 females, with the age of 11.2 (9.2, 13.1) years. Based on the presence of fecaliths, patients were divided into two groups: fecalith group (32 cases) and non-fecalith group (23 cases). No statistically significant differences were observed between the two groups in terms of age, gender, duration of abdominal pain, white blood cell count, neutrophil percentage, diameter of appendix, thickness of appendix clinical symptoms or signs (all P>0.05). The initial treatment success rates were 91% (29/32) in fecalith group and 96% (22/23) in non-fecalith group, with no statistically significant difference ( P=0.632). However, significant differences were noted in stent placement ( χ2=5.85, P=0.026) and procedure time ( Z=4.75, P<0.001). The follow-up duration time was 6.0 (2.0, 12.0) and 7.0 (2.0, 8.5) months for the fecalith and non-fecalith groups, respectively, with no significant difference ( Z=0.05, P=0.962). The recurrence rates were 14% (4/29) in fecalith group and 5% (1/22) in non-fecalith group, with no statistically significant difference ( P=0.375). Conclusions:vERAT can safely and effectively treat pediatric ASA, regardless of the presence or absence of fecaliths. It can provide a new treatment option for ASA.

Objective To evaluate the efficacy of Compound Fipronil Spot-on Solution in repelling canine ticks.Methods A total of 140 dogs infested with ticks were randomly selected from regions in southern and northern China and assigned to four groups:southern test drug group,southern control drug group,northern test drug group,and northern control drug group.Each group comprised 35 dogs.Each dog was administered the prescribed dose.The number of ticks was counted on days 1,7,14,21,and 28 following the administration.The negative conversion and average reduction rates of the tick population were then subjected to statistical analyse.Results The mean efficacy of the test drug was 100%in both the southern and northern cohorts,28 days post-treatment.The control drug showed comparable efficacy,reaching a mean reduction of 100%in both regions by the same time point.No additional clinical manifestations or adverse events were observed across all treated dogs.Conclusions Compound Fipronil Spot-on Solutions effectively treats and prevents ticks in dogs in different regions of China.A single dose remains effective for up to 28 days,thus providing a convenient,effective solution.

Although clinical evidence suggests that nonalcoholic fatty liver disease is an established major risk factor for heart failure, it remains unexplored whether sleep disorder-caused hepatic damage contributes to the development of cardiovascular disease (CVD). Here, our findings revealed that sleep fragmentation (SF) displayed notable hepatic detrimental phenotypes, including steatosis and oxidative damage, along with significant abnormalities in cardiac structure and function. All these pathological changes persisted even after sleep recovery for 2 consecutive weeks or more, displaying memory properties. Mechanistically, persistent higher expression of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in the liver was the key initiator of SF-accelerated damage phenotypes. SF epigenetically controlled the acetylation of histone H3 lysine 27 (H3K27ac) enrichment at the Nox4 promoter and markedly increased Nox4 expression in liver even after sleep recovery. Moreover, fine coordination of the circadian clock and hepatic damage was strictly controlled by BMAL1-dependent Sirtuin 1 (Sirt1) transcription after circadian misalignment. Accordingly, genetic manipulation of liver-specific Nox4 or Sirt1, along with pharmacological intervention targeting NOX4 (GLX351322) or SIRT1 (Resveratrol), could effectively erase the epigenetic modification of Nox4 by reducing the H3K27ac level and ameliorate the progression of liver pathology, thereby counteracting SF-evoked sustained CVD. Collectively, our findings may pave the way for strategies to mitigate myocardial injury from persistent hepatic detrimental memory in diabetic patients.

Objective To explore the molecular mechanism of plasmalemmal vesicle-associated protein(PLVAP)regulating high glucose(HG)induced endocytosis dysfunction in human liver sinusoidal endothelial cells(HLSECs)through Kelch-like epichlorohydrin ECH-associated protein 1(Keap1)/nuclear factor erythroid 2-related factor 2(Nrf2)/Maf protein.Methods HLSECs were cultured in vitro and divided into normal control(NC)group,high glucose(HG)group,PLVAP overexpression recombinant vector(LV-PLVAP)group,lentivirus empty vector(LV-CON)group,HG+LV-PLVAP+Nrf2 inhibitor(ML385)group,HG+LV-PLVAP+Maf inhibitor(Mafenide)group,HG+LV-CON+ML385 group and HG+LV-CON+Mafenide group.Cell activity was detected by CCK-8 assay.The transfection efficiency of LV-PLVAP was observed by fluorescence microscopy.The fluorescence expression of PLVAP and Nrf2 was detected by immunofluorescence.RT-PCR and western blot were used to detect the mRNA and protein expression of PLVAP,Keap1,Nrf2,and Maf,Caveolin-1(CAV-1).Results A lot of green fluorescence appeared in LV-PLVAP and LV-CON groups,however,no green fluorescence was shown in NC group.Immunofluorescence results showed that PLVAP was expressed in cell membrane;Nrf2 was expressed in cytoplasm,and in HG group,Nrf2 was expressed in nucleus.Compared with NC group,the expression of PLVAP,Keap1 mRNA and protein decreased in HG group(P<0.05 or P<0.01),while the expression of Nrf2,Maf and CAV-1 mRNA and protein increased in HG group(P<0.01).Compared with HG group,the expression of PLVAP,Keap1,Nrf2 mRNA and protein increased in HG+LV-PLVAP group(P<0.01),while the expression of Maf,CAV-1 mRNA and protein decreased in HG+LV-PLVAP group(P<0.01).Compared with HG+LV-PLVAP group,the expression of CAV-1 mRNA and protein increased in HG+LV-PLVAP+ML385 and HG+LV-PLVAP+Mafenide groups(P<0.05 or P<0.01).Conclusions PLVAP regulates the expression of CAV-1 through the Keap1/Nrf2/Maf pathway and then regulates HG induced endocytosis dys-function in HLSECs.Over expression of PLVAP alleviates this pathological reaction.

Aim To evaluate the ameliorative effects of different ratios of Renshen-Huangjing(RH) on SPS-induced PTSD-like behaviors in mice,and to investi-gate the action mechanism using bioinformatics analysis and experimental studies.Methods The aqueous ex-tract was extracted in four ratios of RH in a total weight of 60 g,i.e.1∶0(RH1),2∶1(RH2),1∶2(RH3),and 0∶1(RH4).The extraction rates of Rg1,Rb1,and polysaccharides from different ratios of RH were then detected using UPLC-UV method.The SPS model was established,and RH1,RH2,RH3 and RH4(400 mg·kg-1)were administered by intragas-tric gavage for 14 day,followed by behavioral tests to e-valuate the PTSD-like behaviors.The serum CORT,IL-1β and IL-10 were determined by ELISA.The possible targets of action were analyzed using bioinformatics.The expression levels of Calpain-1,PSD95,BDNF and GluN2B in the hippocampus were detected by Western blot.Results The SPS model induced PTSD-like be-haviors in mice.Serum levels of CORT and IL-1β in-creased and level of IL-10 decreased in SPS model.After treatment with different ratios of RHs,RH2 showed the best therapeutic effect,which was manifes-ted in the suppression of PTSD-like behaviors,the re-duction of CORT and IL-1β levels,and the promotion of IL-10 levels;160 overlapping targets might explain the therapeutic effects of RH on PTSD,and these tar-gets were enriched in inhibiting synaptic damage,exer-ting antioxidant properties and suppressing neuroin-flammation,respectively.RH2 prevented the SPS-in-duced decrease in the expression of Calpain-1,PSD95,BDNF and the elevation of GluN2B.Molecular docking showed strong binding of Rg1 and Rb1 to Calpain-1,PSD95,and BDNF,respectively.Conclusions The a-queous extract of RH in a 2∶1 ratio can more effec-tively prevent SPS-induced PTSD-like behaviors,and its effect may be related to targets such as Calpain-1,PSD95,BDNF and GluN2B.

Objective: To explore the diagnosis, treatment and prognosis of primary prostatic signet ring cell carcinoma (SRCC), so as to provide reference for the clinical diagnosis and treatment. Methods: A retrospective analysis was conducted on the clinical data of 6 patients with primary prostatic SRCC treated in Nanjing Drum Tower Hospital during Nov.2020 and Sep.2024.The clinical manifestations, imaging features, treatment methods, histological characteristics and prognosis were summarized. Results: The average age of the patients was (72.00±4.28) years.Varying degrees of dysuria occurred in 4 patients. All patients underwent multi-parametric magnetic resonance imaging (mpMRI) examination before surgery, and the results indicated typical prostate cancer.Preoperative biopsies showed high-grade (Gleason 8-10) prostate acinar adenocarcinoma.Postoperative pathological diagnoses were mixed types of prostate acinar adenocarcinoma and SRCC, and no metastasis was found in the pelvic lymph nodes.All patients were followed up for 1 to 46 months after surgery and are currently alive.Robot-assisted laparoscopic radical prostatectomy only was performed in 3 cases; apalutamide and leuprolide/triptorelin was administered after surgery in 2 cases; bicalutamide + goserelin was administered after surgery in 1 case, who developed bladder metastasis of prostate cancer 24 months later, and the serum prostate-specific antigen (PSA) concentration decreased to a safe level (<0.2 ng/mL) after the use of darolutamide with radiotherapy.No recurrence or metastasis was found in the remaining patients. Conclusion: Primary prostatic SRCC is a rare and highly aggressive malignant tumor of the prostate.The diagnosis depends on pathological examinations due to lack of specific imaging features and clinical manifestations.The prognosis is poor, and there is currently no standardized treatment.The combined use of surgery, hormonotherapy and radiotherapy can help improve the survival rate of patients.

Continuous manufacturing, as an innovative pharmaceutical production model, offers advantages such as high production efficiency and ease of control compared to traditional batch production, aligning with the future trend of drug production moving toward greater efficiency and intelligence. However, the development of continuous manufacturing technology in wet granulation has been slow. On one hand, this is closely related to its high technical complexity, substantial equipment investment costs, and stringent process control requirements. On the other hand, the long-term use of the traditional batch production model has created strong path dependence, and the lack of mature standardized processes further increases the difficulty of technological transformation. To promote the deep integration of wet granulation technology with continuous manufacturing, this review systematically outlines the current application of wet granulation in continuous manufacturing. It focuses on the development of key technologies such as online detection, process modeling, and process scale-up, with the aim of providing a reference for process innovation and application in wet granulation.
Drug Compounding/instrumentation* ; Technology, Pharmaceutical/methods* ; Drugs, Chinese Herbal/chemistry* ; Models, Theoretical

AIM To establish the near-infrared spectroscopy quantitative models for moisture,23-acetylalismol B and 23-acetylalismol C in Alismatis Rhizoma decoction pieces.METHODS The near-infrared spectroscopy(NIRS)data were collected in 95 batches of decoction pieces,after which drying method was adopted in the content determination of moisture,HPLC was applied to determining the contents of 23-acetylalismol B and 23-acetylalismol C,the quantitative models were established by partial least squares method combined with feature extraction algorithms.RESULTS The model training determination coefficients were 0.952 6,0.958 1 and 0.920 8,along with the prediction determination coefficients of 0.930 0,0.905 2 and 0.906 4,the residual prediction deviations(PRD)of 4.00,3.58 and 3.46,and the root mean square error ratios of prediction values to calibration values(RMSEP/RMSEC)of 1.15,1.11 and 1.06,respectively.CONCLUSION The quantitative models based on NIRS exhibit good prediction effects,which can be used for the rapid quality detection of Alismatis Rhizoma decoction pieces.