OBJECTIVE To establish a population pharmacokinetic(PopPK)model to predict the PK characteristics of GLS-010 in humans.METHODS Fifty-eight cynomolgus monkeys were used,18 of which were randomly divided into three groups and received a single intravenous infusion of GLS-010 at doses of 2,6,and 18 mg·kg-1,respectively.The rest were randomly assigned to four groups and received multiple intravenous infusions of GLS-010 at doses of 0,5,25,and 100 mg·kg-1,respectively,once a week(quaque week,qw)for five consecutive weeks.Blood samples were collected before and after administration.The concentrations of GLS-010 in the monkey serum were measured using a validated enzyme-linked immunosorbent assay,while those of anti-drug antibodies(ADA)in the cynomolgus monkey serum were determined by ultra-sensitive electrochemiluminescence immunoassay.The PK data on GLS-010 in cynomolgus monkeys was obtained,and the drug-time curves were plotted.A PopPK model was constructed using non-compartmental analysis and evaluated by goodness-of-fit plots and visual predictive checks.The constructed PopPK model was used to predict the PK characteristics in humans,which were finally compared with actual Phase Ⅰ clinical study results for validation.RESULTS The predictive results of the PopPK model were highly consistent with the actual Phase Ⅰ clinical study results.The model was able to predict the human PK characteristics under various dosing regimens,including 1 mg·kg-1 quaque 2 weeks(q2w),4 mg·kg-1(q2w),240 mg(q2w),240 mg(q3w),and 10 mg·kg-1(q2w).The predicted maximum plasma concentrations(Cmax)were 24.8,99.1,85.0,85.0,and 247.8 mg·L-1,respectively,and the AUC0-336h was 4 902.0,20 060.0,17 147.7,22 145.7(AUC0-504h),and 50 817.6 mg·h·L-1,respectively.The safety risks for the corresponding dosing regimens were 47.3,11.6,13.5,10.5,and 4.6,respectively.The predicted receptor occupancy at steady state(ROss)at Cmax,average plasma concentration(Cavg),and minimum plasma concentration(Cmin)were 38.8％,72.7％,69.4％,64.1％and 87.2％,29.1％,63.8％,60.0％,49.8％and 82.1％,21.9％,55.5％,51.3％,36.3％and 76.7％,respectively.CONCLUSION The PopPK model can effectively predict the human PK characteristics under different dosing regimens with high consistency with actual Phase Ⅰ clinical study results,which can serve as an important reference for selection of safe and effective doses for first-in-human research.

The receptor-binding region(RBD)of the spike protein(S)on the surface of porcine epi-demic diarrhea virus(PEDV)is a critical structural domain mediating viral invasion of host cells and serves as a key target for inducing neutralizing antibodies.In order to prepare antibodies that can be used to study the biological function of PEDV RBD and develop novel diagnostic and thera-peutic methods,recombinant RBD protein expressed in Sf9 insect cells was utilized as an immuno-gen to immunize BALB/c mice.Monoclonal antibodies(mAbs)were generated via hybridoma tech-nology,and positive hybridoma clones were screened using indirect ELISA.The reactivity of the mAbs was subsequently characterized.The results of ELISA,Western blot,and indirect immuno-fluorescence assay(IFA)showed that three monoclonal antibodies screened(3E5,4F9 and 5A5)had good reactivity with the virus and RBD protein.Antibody subtype results showed that 3E5 and 4F9 were of IgG1 subtypes and 5A5 was of IgM subtype.Neutralization assay further revealed that 3E5 monoclonal antibody had viral neutralizing activity.In this study,three monoclonal antibodies against PEDV RBD proteins were successfully prepared,providing the basis for the study of the bi-ological function of RBD proteins,PEDV serologic detection and vaccine development.

The receptor-binding region(RBD)of the spike protein(S)on the surface of porcine epi-demic diarrhea virus(PEDV)is a critical structural domain mediating viral invasion of host cells and serves as a key target for inducing neutralizing antibodies.In order to prepare antibodies that can be used to study the biological function of PEDV RBD and develop novel diagnostic and thera-peutic methods,recombinant RBD protein expressed in Sf9 insect cells was utilized as an immuno-gen to immunize BALB/c mice.Monoclonal antibodies(mAbs)were generated via hybridoma tech-nology,and positive hybridoma clones were screened using indirect ELISA.The reactivity of the mAbs was subsequently characterized.The results of ELISA,Western blot,and indirect immuno-fluorescence assay(IFA)showed that three monoclonal antibodies screened(3E5,4F9 and 5A5)had good reactivity with the virus and RBD protein.Antibody subtype results showed that 3E5 and 4F9 were of IgG1 subtypes and 5A5 was of IgM subtype.Neutralization assay further revealed that 3E5 monoclonal antibody had viral neutralizing activity.In this study,three monoclonal antibodies against PEDV RBD proteins were successfully prepared,providing the basis for the study of the bi-ological function of RBD proteins,PEDV serologic detection and vaccine development.

Objective To analyze the expression of serum mannose binding lectin associated serine protease 1(MASP1)and protein kinase D2(PRKD2)in locally advanced cervical cancer(LACC)patients,and their relationship with the efficacy of neoadjuvant chemotherapy(NACT).Methods The clinical data of 98 LACC patients(LACC group)treated with NACT in the hospital from March 2019 to April 2021 were retrospective-ly selected,and they were divided into effective group(n=76)and ineffective group(n=22)according to the efficacy.Fifty patients with early cervical cancer diagnosed and treated in the same period were selected as the early cervical cancer group,and 50 female health examination subjects were selected as the healthy control group.Enzyme-linked immunosorbent assay was applied to detect serum levels of MASP1 and PRKD2.Logis-tic regression analysis was applied to analyze the influencing factors of NACT efficacy.The evaluation value of serum MASP1 and PRKD2 on the efficacy of NACT were analyzed by the receiver operating characteristic curve.Results The serum levels of MASP1 and PRKD2 in the LACC group were higher than those in the ear-ly cervical cancer group and the control group,and the differences were statistically significant(P＜0.05).The serum levels of MASP1 and PRKD2 in LACC group were correlated with International Federation of Gy-necology and Obstetrics(FIGO)stage and pathological grade,and the serum levels of MASP1 and PRKD2 in patients with FIGO stage Ⅲ and pathological grade G3 were higher(P＜0.05).FIGO stage Ⅲ proportion,se-rum MASP1 and serum PRKD2 levels in NACT ineffective group were higher than those in effective group(P＜0.05).FIGO stage Ⅲ,serum MASP1 and serum PRKD2 were independent risk factors for NACT efficacy in LACC patients(P＜0.05).The area under the curve of serum MASP1 and PRKD2 combined to evaluate the efficacy of NACT was 0.883(95％CI:0.828-0.935),which was larger than 0.802(95％CI:0.761-0.846)and 0.825(95％CI:0.764-0.852)predicted by the single index,and the difference were statistically significant(Z=4.111,5.012,both P＜0.001).Conclusion Level of serum MASP1 and PRKD2 in LACC pa-tients are increase,which are independent risk factors affecting the efficacy of NACT.Combined detection of MASP1 and PRKD2 has high predictive value for NACT efficacy.

OBJECTIVE To establish a population pharmacokinetic(PopPK)model to predict the PK characteristics of GLS-010 in humans.METHODS Fifty-eight cynomolgus monkeys were used,18 of which were randomly divided into three groups and received a single intravenous infusion of GLS-010 at doses of 2,6,and 18 mg·kg-1,respectively.The rest were randomly assigned to four groups and received multiple intravenous infusions of GLS-010 at doses of 0,5,25,and 100 mg·kg-1,respectively,once a week(quaque week,qw)for five consecutive weeks.Blood samples were collected before and after administration.The concentrations of GLS-010 in the monkey serum were measured using a validated enzyme-linked immunosorbent assay,while those of anti-drug antibodies(ADA)in the cynomolgus monkey serum were determined by ultra-sensitive electrochemiluminescence immunoassay.The PK data on GLS-010 in cynomolgus monkeys was obtained,and the drug-time curves were plotted.A PopPK model was constructed using non-compartmental analysis and evaluated by goodness-of-fit plots and visual predictive checks.The constructed PopPK model was used to predict the PK characteristics in humans,which were finally compared with actual Phase Ⅰ clinical study results for validation.RESULTS The predictive results of the PopPK model were highly consistent with the actual Phase Ⅰ clinical study results.The model was able to predict the human PK characteristics under various dosing regimens,including 1 mg·kg-1 quaque 2 weeks(q2w),4 mg·kg-1(q2w),240 mg(q2w),240 mg(q3w),and 10 mg·kg-1(q2w).The predicted maximum plasma concentrations(Cmax)were 24.8,99.1,85.0,85.0,and 247.8 mg·L-1,respectively,and the AUC0-336h was 4 902.0,20 060.0,17 147.7,22 145.7(AUC0-504h),and 50 817.6 mg·h·L-1,respectively.The safety risks for the corresponding dosing regimens were 47.3,11.6,13.5,10.5,and 4.6,respectively.The predicted receptor occupancy at steady state(ROss)at Cmax,average plasma concentration(Cavg),and minimum plasma concentration(Cmin)were 38.8％,72.7％,69.4％,64.1％and 87.2％,29.1％,63.8％,60.0％,49.8％and 82.1％,21.9％,55.5％,51.3％,36.3％and 76.7％,respectively.CONCLUSION The PopPK model can effectively predict the human PK characteristics under different dosing regimens with high consistency with actual Phase Ⅰ clinical study results,which can serve as an important reference for selection of safe and effective doses for first-in-human research.

Premature ovarian insufficiency(POI),also known as"ovarian insufficiency",has an incidence of 1％～5％.The incidence has been on the rise in recent years,seriously affecting women's physical and mental health and quality of life.At present,the cause and mechanisms of POI are still unclear,and the method and applications of model construction are also confusing.Most models have some shortcomings in pertinence and stability.The limitations greatly limit research into the clinical diagnosis and treatment of POI.This paper summarizes and discusses the etiology and pathogenesis of POI and the construction of POI animal models to provide a comprehensive reference for those studying POI.

Porcine deltacoronavirus(PDCoV),a newly discovered virus in recent years,can cause severe diarrhea,vomiting,dehydration and even death in piglets.S protein is an important structur-al protein of PDCoV,which determines the host or tissue tropism of the virus,and is an important target for the development of PDCoV vaccines and detection methods.In order to prepare mono-clonal antibody(MAb)against the S protein of PDCoV,the recombinant plasmid of S protein was constructed based on the extracellular domain sequence of S protein of PDCoV epidemic strain in China and transformed into DH10Bac competent cells.The recombinant bacmid was identified by blue-white spot screening and PCR.BALB/c mice were immunized with S protein,and the spleen cells of immunized mice were fused with myeloma cells.The positive hybridoma cells that secreted stable antibodies were screened by indirect ELISA and subcloning.Five hybridoma cell superna-tants(MAb)specifically recognizing S protein(2E5,4D5,5D10,2F7 and 5A9)were identified by Western blot and immunofluorescence assay(IFA).Subsequently,the neutralization test showed that three of the monoclonal antibodies(2E5,4D5 and 5D10)could neutralize the virus to varying degrees.The S protein was successfully expressed and 5 monoclonal antibodies that can stably se-crete and specifically bind to PDCoV and S protein were prepared,which laid an important founda-tion for further research on the structure and function of S protein,the development of detection methods for PDCoV infection,and the prevention or treatment of PDCoV infection.

Objective To analyze the predictive value of serum micro RNA(miR)-411-5p and micro RNA(miR)-485-5p levels in preeclampsia(PE)pregnant women combined with ultrasound blood flow indicators for perinatal outcomes.Methods A total of 88 pregnant women with PE who were enrolled in the obstetric card and underwent cesarean section in Beijing Nuclear Industry Hospital from January 2020 to December 2021 were selected as the study subjects(PE group).In addition,90 normal pregnant women who delivered by cesarean section due to other reasons in the same period were regarded as the control group.The differences of pulsitility index(PI)and resistance index(RI)were compared between the two groups.The serum levels of miR-411-5p and miR-485-5p were measured and compared between the two groups by real time fluorescent quantitative PCR(qRT-PCR).According to the different perinatal outcomes of PE patients,they were grouped into good outcome group and poor outcome group.The differences of PI,RI,miR-411-5p and miR-485-5p between the two groups were compared.The predictive effect of PI,RI,miR-411-5p,miR-485-5p and combined detection on the adverse perinatal outcome of PE patients was analyzed by the receiver operating characteristic(ROC)curve method.Results The RI(0.79±0.08)and PI values(1.82±0.08)of pregnant women in the PE group were higher than those in the control group(0.66±0.06,1.38±0.15),the serum levels of miR-411-5p(0.32±0.09)and miR-485-5p(0.26±0.03)were lower than those in the control group(1.01±0.08,1.02±0.09),and the differences were statistically significant(t=12.283,24.339,54.091,75.231,all P＜0.001).The RI(0.83±0.08)and PI(1.86±0.09)values of PE patients in the poor outcome group were higher than those in the good outcome group(0.70±0.07,1.71±0.07),the serum levels of miR-411-5p(0.27±0.02)and miR-485-5p(0.24±0.02)were lower than those in the good outcome group(0.45±0.04,0.31±0.04),and the differences were statistically significant(t=11.545,12.428,37.840,14.716,all P＜0.001).The areas under the curve of ultrasound blood flow index RI and PI to predict the adverse perinatal outcome of PE patients were 0.838(sensitivity was 90.8％,specificity was 65.2％),and 0.758(sensitivity was 50.8％,specificity was 91.3％),respectively.The areas under the curve of serum miR-411-5p and miR-485-5p in predicting adverse perinatal outcome of PE patients were 0.830(sensitivity was 90.8％,specificity was 73.9％),and 0.769(sensitivity was 95.4％,specificity was 61.9％),respectively.The area under the curve of the four combined tests to predict the adverse perinatal outcome of PE patients was 0.976(sensitivity was 98.5％,specificity was 91.3％).Conclusion The levels of miR-411-5p and miR-485-5p in serum of PE pregnant women were decreased,and the combination of miR-411-5p and miR-485-5p and ultrasound blood flow indicators PI and RI may predict the perinatal fetal outcome of PE pregnant women.

Background and purpose:Radical surgery after neoadjuvant immunotherapy combined with chemotherapy(nICT)in patients with locally advanced esophageal squamous cell carcinoma(LAESCC)has good efficacy and safety,and it can improve the patients'pathological complete remission(pCR)rate,main pathologic response(MPR)rate and R0 resection rate.The prognosis of patients with pCR/MPR after nICT is significantly better compared with patients without pCR.The prognosis of patients achieving pCR/MPR after neoadjuvant therapy has been demonstrated to be significantly better than that of patients with non-pCR/MPR.Therefore,finding predictive factors of pCR/MPR is beneficial for us to screen out the advantageous populations for combination therapy.The aim of this study was to investigate the value of clinical data of patients with LAESCC before and after nICT in predicting the degree of remission of different pathologies after radical surgery following neoadjuvant treatment and to observe the safety of the treatment.Methods:Data of patients with locally LAESCC who underwent radical surgery after nICT from January 2019 to June 2023 at the Affiliated Hospital of Chuanbei Medical College were collected.The clinical data of all patients as well as some blood,inflammation and nutritional indexes of patients before and after neoadjuvant therapy were collected,and the patients were grouped according to the different degrees of pathological remission after neoadjuvant therapy.Data were analyzed by multi-group comparative analysis of variance(ANOVA)and LSD-t post-hoc test.We explored the factors that had an influence on the different degrees of pathological remission,and collected and recorded the patients'adverse reactions during neoadjuvant therapy as well as their eventual surgeries.Results:Data of 62 patients with LAESCC treated with nICT who underwent radical surgery were collected.Only one patient showed grade 4 myelosuppression during neoadjuvant therapy,and the rest of the patients had adverse reactions≤grade 2.The R0 resection rate of the surgery was 98.39%.The present study was compared with the previous studies of LAESCC treated with neoadjuvant chemotherapy followed by radical surgery performed in Affiliated Hospital of Chuanbei Medical College.Compared with the previous studies conducted in our center,no significant difference was observed in terms of operation time,intraoperative bleeding,postoperative hospitalization time and surgical complications.The postoperative pathologic results showed that the pCR rate was 22.58%(14/62),and the MPR rate was 40.32%(25/62).According to the different tumor regression grade(TRG)after surgery,patients were divided into 3 groups of TRG1,TRG2 and TRG3-4,and differences in the platelet distribution width(PDW)before neoadjuvant therapy and the preoperative neutrophil-to-lymphocyte ratio(NLR)after neoadjuvant therapy were statistically significant among the 3 groups(P＜0.05).Further intra-group two-by-two comparisons of PDW before neoadjuvant therapy and NLR before surgery after neoadjuvant therapy were performed for the three groups of patients,respectively,and it was found that the PDW and NLR in the TRG2 group were lower compared with the TRG3-4 group,and the differences were statistically significant(P＜0.05).Conclusion:Radical surgery after nICT treatment in patients with LAESCC can have high R0 resection rate,pCR rate,MPR rate and reliable safety,and the lower PDW of patients before neoadjuvant therapy and the lower NLR of patients before surgery after neoadjuvant therapy predict better pathological remission efficacy.

OBJECTIVE To evaluate the mechanisms underlying the antidepressant effect of ZBH2012001,a novel serotonin and norepinephrine reuptake inhibitor(SNRI),in general and its ability to enhance monoaminergic transmission and suppress neuroinflammation in particular.METHODS① Male ICR mice were divided into vehicle(distilled water),duloxetine(DLX,10 or 20 mg·kg-1)and ZBH2012001(5,10 and 20 mg·kg-1)groups.One hour following ig administration,the antidepressant effect of ZBH2012001 was evaluated using the tail suspension test(TST)and forced swimming test(FST).② Radioligand binding assay was conducted to evaluate the affinity of ZBH2012001 for human serotonin transporters(hSERTs)and human norepinephrine transporters(hNETs).③ Mice were divided into vehicle(distilled water),DLX(10 or 20 mg·kg-1)and ZBH2012001(5,10 and 20 mg·kg-1)groups.One hour following drug administration,the 5-hydroxytryptophan(5-HTP)-induced head-twitch test or yohimbine-induced lethality test were performed to evaluate the effect of ZBH2012001 on the function of the 5-hydroxytryptamine(5-HT)and norepinephrine(NE)systems.④ Mice were divided into vehicle(distilled water+0.1%acetic acid),reserpine model(distilled water+reserpine 5 mg·kg-1),DLX(DLX 20 mg·kg-1+reserpine 5 mg·kg-1)and ZBH2012001(ZBH2012001 5,10 and 20 mg·kg-1+reserpine 5 mg·kg-1)groups.One hour following drug administration,reserpine was injected intraperitoneally to establish a monoamine-depletion model.The ptosis,akinesia,and hypothermia assays were performed to evaluate the effect of ZBH2012001 on the down-regulation of the reserpine-induced monoamine system.The TST in mice was used to evaluate the effect of ZBH2012001 on reserpine-induced depressive-like behavior while high-performance liquid chromatography with electrochemical detection(HPLC-ECD)was used to measure the levels of monoamines and their metabolites in the hippocampal tissue of reserpine-induced monoamine-depletion mice.ELISA was employed to detect the contents of tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in the hippocampal tissue of reserpine-induced monoamine-depletion mice.Western blotting was used to assess the expressions of ionized calcium-binding adapter molecule-1(Iba-1)and nuclear factor-kappa B(NF-κB)in the hippocampal tissue of reserpine-induced monoamine-depletion mice.RESULTS ① Compared with the vehicle group,ZBH2012001(5,10 and 20 mg·kg-1)significantly reduced the immobility time both in the TST in mice(P<0.01,respectively),and ZBH2012001(20 mg·kg-1)and in the FST in mice(P<0.05).② ZBH2012001 competitively inhibited the binding of[3H]-imipramine to hSERTs and[3H]-nisoxetine to hNETs,with the half maximal inhibitory concentration(IC50)values of 84.95 and 712.90 nmol·L-1,respectively.③Com-pared with the vehicle group,ZBH2012001(10 and 20 mg·kg-1)significantly increased the head twitches induced by 5-HTP in mice(P<0.01,respectively)and increased the mortality rate in mice induced by yohimbine(P<0.05,P<0.01).④ In the reserpine-induced monoamine-depletion model in mice,compared with the vehicle group,mice in the reserpine model group exhibited ptosis,akinesia and hypothermia feature(P<0.01,respectively),significantly prolonged immobility time in the TST(P<0.01),significantly decreased the levels of NE,5-HT and dopamine(DA)(P<0.05,P<0.01),significantly increased the metabolic conversion rate of 5-HT and DA(P<0.01,respectively),significantly elevated levels of TNF-α and IL-6(P<0.05,respectively),and significantly increased expressions of Iba-1 and NF-κB(P<0.05,respectively)in the hippocampus.Compared with the model group,ZBH2012001(5,10 and 20 mg·kg-1)significantly antagonized ptosis and hypothermia behaviors induced by reserpine(P<0.01,respectively),ZBH2012001(10 and 20 mg·kg-1)significantly shortened the immobility time in reserpine-treated mice(P<0.05,P<0.01),ZBH2012001(20 mg·kg-1)significantly increased the levels of NE and 5-HT in the hippocampus of reserpine-treated mice(P<0.05,respectively),decreased the metabolic conversion rate of 5-HT(P<0.05),significantly reduced the contents of TNF-α and IL-6 in the hippocampus of reserpine-treated mice(P<0.05,respectively),ZBH2012001(5,10 and 20 mg·kg-1)significantly reduced the expression of Iba-1 protein in the hippocampus of reserpine-treated mice(P<0.01,respec-tively),and ZBH2012001(20 mg·kg-1)significantly reduced the expression of NF-κB protein in the hippocampus of reserpine-treated mice(P<0.05).CONCLUSION ZBH2012001 exerts its antidepres-sant effect through a dual mechanism involving monoamine enhancement and inflammation suppres-sion.