Neonatal diabetes mellitus （NDM） is a rare monogenic disorder primarily caused by insufficient insulin secretion resulting from mutations in the KCNJ11 and ABCC8 genes. Sulfonylureas， represented by glibenclamide， have become the standard therapy for this type of NDM by precisely closing the mutated ATP-sensitive potassium channels in pancreatic β cells， thereby restoring insulin secretion. Clinical studies confirm that sulfonylureas enable over 90% of patients to successfully transition from insulin to oral treatment， achieving long-term stable glycemic control and improving neurological outcomes to a certain extent. In terms of safety， severe hypoglycemia induced by sulfonylureas is relatively rare and gastrointestinal reactions are mild； moreover， sulfonylureas show good long-term tolerability， and have no adverse effects on child growth and development. In the future， by further refining the full-chain management pathway of “rapid genetic diagnosis-early intervention-specialized dosage forms-long-term follow-up”， the clinical application of sulfonylureas is expected to provide NDM patients with an optimized treatment regimen and maximize their health benefits.

Objective:To explore the values of reticulin fiber staining (RFS) in evaluating bone marrow (BM) involvement of lymphoma and in grading of BM biopsy from adult lymphoma patients.Methods:Retrospectively,354 cases of adult lymphoma were collected from November 2023 to May 2024 at Peking University Cancer Hospital. BM samples were stained with RFS and immunohistochemical staining (IHC), and flow cytometry (FCM) was also performed with the BM aspirations simultaneously. RFS was graded according to the European Consensus, as high grade (grade 2-3) indicating BM involvement in the study. BM involvement was considered as definite if no less than two positive findings among IHC, FCM, and RFS. Statistical analyses were performed via SPSS software (V23.0).Results:In this series, 52.3% (185/354) of the patients were male; 35.0% (124/354) aged >60 years; BM involvements were found in 34.5% (122/354) cases with high grade of RFS, which, in turn, were lymphoblastic leukemia/lymphoma (ALL/LBL) group (4/4), indolent B-cell lymphoma (IndBCL) group (49.1%, 53/108), transformed B-cell lymphoma (TrBCL) group (2/5), invasive B-cell lymphoma (InvBCL) group (26.5%, 41/155), T and NK cell lymphoma (TNKCL) group (27.3%, 12/44) and classical Hodgkin lymphoma (CHL) group (26.3%, 10/38); if classified by specific types, T-ALL/LBL (2/2), B-ALL/LBL (2/2) and CLL/SLL (8/10) ranked top three. In terms of the positive rate of BM involvement evaluated by RFS, no significant difference was seen between either gender or age groups ( χ2=3.416, P=0.332 and χ2=4.200, P=0.241); however, significant differences were observed between different lymphoma groups and types ( χ2=29.961, P=0.012 and χ2=102.546, P<0.001, respectively). BM invasion rates indicated by IHC and FCM were 25.4% (90/354) and 13.8% (49/354), respectively. The overall BM invasion rate was 24.3% (86/354), and the sensitivity of RFS, IHC, and FCM was 90.8%, 97.8%, and 55.8%, and specificity was 84.1%, 99.6%, and 98.9%, respectively. Overall, the concordance rate of RFS with IHC and FCM was 83.6% and 74.0%, respectively, including 85.8% and 74.2% for InvBCL group, 79.6% and 75.0% for IndBCL group, 84.1% and 75.0% for TNKCL group, 81.6% and 73.7% for CHL group, 5/5 and 2/5 for TrBCL group, and 4/4 and 3/4 for ALL/LBL group. Conclusions:In the evaluation of BM involvement status of adult lymphoma, high sensitivity and specificity are observed by RFS, and high concordance is also noted with both IHC and FCM. Thus, the BM infiltrating status of adult lymphoma could be evaluated more accurately by a combined usage of the three methods.

Objective:To explore the values of reticulin fiber staining (RFS) in evaluating bone marrow (BM) involvement of lymphoma and in grading of BM biopsy from adult lymphoma patients.Methods:Retrospectively,354 cases of adult lymphoma were collected from November 2023 to May 2024 at Peking University Cancer Hospital. BM samples were stained with RFS and immunohistochemical staining (IHC), and flow cytometry (FCM) was also performed with the BM aspirations simultaneously. RFS was graded according to the European Consensus, as high grade (grade 2-3) indicating BM involvement in the study. BM involvement was considered as definite if no less than two positive findings among IHC, FCM, and RFS. Statistical analyses were performed via SPSS software (V23.0).Results:In this series, 52.3% (185/354) of the patients were male; 35.0% (124/354) aged >60 years; BM involvements were found in 34.5% (122/354) cases with high grade of RFS, which, in turn, were lymphoblastic leukemia/lymphoma (ALL/LBL) group (4/4), indolent B-cell lymphoma (IndBCL) group (49.1%, 53/108), transformed B-cell lymphoma (TrBCL) group (2/5), invasive B-cell lymphoma (InvBCL) group (26.5%, 41/155), T and NK cell lymphoma (TNKCL) group (27.3%, 12/44) and classical Hodgkin lymphoma (CHL) group (26.3%, 10/38); if classified by specific types, T-ALL/LBL (2/2), B-ALL/LBL (2/2) and CLL/SLL (8/10) ranked top three. In terms of the positive rate of BM involvement evaluated by RFS, no significant difference was seen between either gender or age groups ( χ2=3.416, P=0.332 and χ2=4.200, P=0.241); however, significant differences were observed between different lymphoma groups and types ( χ2=29.961, P=0.012 and χ2=102.546, P<0.001, respectively). BM invasion rates indicated by IHC and FCM were 25.4% (90/354) and 13.8% (49/354), respectively. The overall BM invasion rate was 24.3% (86/354), and the sensitivity of RFS, IHC, and FCM was 90.8%, 97.8%, and 55.8%, and specificity was 84.1%, 99.6%, and 98.9%, respectively. Overall, the concordance rate of RFS with IHC and FCM was 83.6% and 74.0%, respectively, including 85.8% and 74.2% for InvBCL group, 79.6% and 75.0% for IndBCL group, 84.1% and 75.0% for TNKCL group, 81.6% and 73.7% for CHL group, 5/5 and 2/5 for TrBCL group, and 4/4 and 3/4 for ALL/LBL group. Conclusions:In the evaluation of BM involvement status of adult lymphoma, high sensitivity and specificity are observed by RFS, and high concordance is also noted with both IHC and FCM. Thus, the BM infiltrating status of adult lymphoma could be evaluated more accurately by a combined usage of the three methods.

Background: Peste des petits ruminants (PPR), caused by the PPR virus (PPRV), is an acute and fatal contagious disease that mainly infects goats, sheep, and other artiodactyls.Peripheral blood mononuclear cells (PBMCs) are considered the primary innate immune cells. Objectives: PBMCs derived from goats were infected with PPRV and analyzed to detect the relationship between PPRV replication and apoptosis or the inflammatory response. Methods: Quantitative real-time polymerase chain reaction was used to identify PPRV replication and cytokines expression. Flow cytometry was conducted to detect apoptosis and the differentiation of CD4+ and CD8+T cells after PPRV infection. Results: PPRV stimulated the differentiation of CD4+ and CD8+ T cells. In addition, PPRV induced apoptosis in goat PBMCs. Furthermore, apoptosis and the inflammatory response induced by PPRV could be suppressed by Z-VAD-FMK and Z-YVAD-FMK, respectively.Moreover, the virus titer of PPRV was attenuated by inhibiting caspase-1-dependent apoptosis and inflammation. Conclusions This study showed that apoptosis and the inflammatory response play an essential role in PPR viral replication in vitro, providing a new mechanism related to the cell host response.

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases.The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions The spbELISA has practical applications in assessing the vaccination status of large pig herds.

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Singledomain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Singledomain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells (SGC-7901). Methods Different concentrations of coix seed oil (2, 4, 8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation, and flow cytometry for drug-induced cell apoptosis, scratch test for cell migration inhibited by coix seed oil, Transwell chamber for drug-inhibited cell invasion, and Western blot for expression of related proteins PRMT5, PI3 K and AKT. Results The results of MTT showed that 2, 4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells, and the inhibition rate of 2 mg/mL was (30. 02 ± 1. 56) ％, which showed significant difference compared to the control group (P ＜ 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were (16. 25 ± 2. 54) ％, (12. 60 ± 1. 12) ％, respectively, and which showed a significant difference compared with (2. 0 ± 1. 22) ％ in the control group (P ＜ 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2, 4 mg/mL of coix seed oil had significant difference when compared to control group (P ＜0. 01). The results of invasion experiments showed that 2, 4 mg/mL of coix seed oil could significantly inhibit cell invasion, and the number of cell invasion was (134. 00 ± 2. 86), (167. 00 ±0. 99), respectively, which showed significant difference compared to (268. 00 ± 2. 05) in the control group (P ＜ 0. 01). The migration number in 8 mg/mL coix seed oil group was (167 ± 0. 99), a significant difference was observed when compared to the control group (P ＜ 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5, PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation, migration and invasion of gastric cancer SGC-7901 cells, its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules, induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.

Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells (SGC-7901). Methods Different concentrations of coix seed oil (2, 4, 8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation, and flow cytometry for drug-induced cell apoptosis, scratch test for cell migration inhibited by coix seed oil, Transwell chamber for drug-inhibited cell invasion, and Western blot for expression of related proteins PRMT5, PI3 K and AKT. Results The results of MTT showed that 2, 4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells, and the inhibition rate of 2 mg/mL was (30. 02 ± 1. 56) ％, which showed significant difference compared to the control group (P ＜ 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were (16. 25 ± 2. 54) ％, (12. 60 ± 1. 12) ％, respectively, and which showed a significant difference compared with (2. 0 ± 1. 22) ％ in the control group (P ＜ 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2, 4 mg/mL of coix seed oil had significant difference when compared to control group (P ＜0. 01). The results of invasion experiments showed that 2, 4 mg/mL of coix seed oil could significantly inhibit cell invasion, and the number of cell invasion was (134. 00 ± 2. 86), (167. 00 ±0. 99), respectively, which showed significant difference compared to (268. 00 ± 2. 05) in the control group (P ＜ 0. 01). The migration number in 8 mg/mL coix seed oil group was (167 ± 0. 99), a significant difference was observed when compared to the control group (P ＜ 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5, PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation, migration and invasion of gastric cancer SGC-7901 cells, its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules, induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.

Objective To identify the influence of chemotherapy-induced serum alkaline phosphatase (ALP) elevation on the tumor-free survival (TFS) in patients of gastric carcinoma after radical gastrectomy.Methods The clinical data of 189 gastric carcinoma patients receiving radical surgery and postoperative adjuvant chemotherapy between Jan,2010 and Dec,2015 were reviewed and statistically analyzed.Results The TFS of patients with serum ALP elevation was obviously inferior than those without ALP elevation (x2 =5.717,P =0.017),serum ALP elevation was an independent risk factor influencing patients' TFS (HR =2.178,P =0.032),the degree of serum ALP elevation was associated with patients' TFS (x2 =4.627,P =0.031).Conclusion Serum ALP elevation during postoperative chemotherapy indicates the increases of recurrence or metastasis rate of gastric cancer patients after radical gastrectomy.