Objective To explore the mechanism by which soybean isoflavone(SI)reduces calcium overload induced by cerebral ischemia-reperfusion(I/R).Methods Forty-eight SD rats were randomized into 4 groups to receive sham operation,cerebral middle artery occlusion for 2 h followed by 24 h of reperfusion(I/R model group),or injection of adeno-associated virus carrying Frizzled-2 siRNA or empty viral vector into the lateral cerebral ventricle after modeling.Western blotting was used to examine Frizzled-2 knockdown efficiency and changes in protein expressions in the Wnt/Ca2+signaling pathway.Calcium levels and pathological changes in the ischemic penumbra(IP)were measured using calcium chromogenic assay and HE staining,respectively.Another 72 SD randomly allocated for sham operation,I/R modeling,or soy isoflavones pretreatment before modeling were examined for regional cerebral blood flow using a Doppler flowmeter,and the cerebral infarct volume was assessed using TTC staining.Pathologies in the IP area were evaluated using HE and Nissl staining,and ROS level,Ca2+level,cell apoptosis,and intracellular calcium concentration were analyzed using immunofluorescence assay or flow cytometry;the protein expressions of Wnt5a,Frizzled-2,and P-CaMK II in the IP were detected with Western blotting and immunohistochemistry.Results In rats with cerebral I/R,Frizzled-2 knockdown significantly lowered calcium concentration(P<0.001)and the expression levels of Wnt5a,Frizzled-2,and P-CaMK II in the IP area.In soy isoflavones-pretreated rats,calcium concentration,ROS and MDA levels,cell apoptosis rate,cerebral infarct volume,and expression levels of Wnt/Ca2+signaling pathway-related proteins were all significantly lower while SOD level was higher than those in rats in I/R model group.Conclusion Soy isoflavones can mitigate calcium overload in rats with cerebral I/R by inhibiting the Wnt/Ca2+signaling pathway.

Objective To explore the mechanism by which soybean isoflavone(SI)reduces calcium overload induced by cerebral ischemia-reperfusion(I/R).Methods Forty-eight SD rats were randomized into 4 groups to receive sham operation,cerebral middle artery occlusion for 2 h followed by 24 h of reperfusion(I/R model group),or injection of adeno-associated virus carrying Frizzled-2 siRNA or empty viral vector into the lateral cerebral ventricle after modeling.Western blotting was used to examine Frizzled-2 knockdown efficiency and changes in protein expressions in the Wnt/Ca2+signaling pathway.Calcium levels and pathological changes in the ischemic penumbra(IP)were measured using calcium chromogenic assay and HE staining,respectively.Another 72 SD randomly allocated for sham operation,I/R modeling,or soy isoflavones pretreatment before modeling were examined for regional cerebral blood flow using a Doppler flowmeter,and the cerebral infarct volume was assessed using TTC staining.Pathologies in the IP area were evaluated using HE and Nissl staining,and ROS level,Ca2+level,cell apoptosis,and intracellular calcium concentration were analyzed using immunofluorescence assay or flow cytometry;the protein expressions of Wnt5a,Frizzled-2,and P-CaMK II in the IP were detected with Western blotting and immunohistochemistry.Results In rats with cerebral I/R,Frizzled-2 knockdown significantly lowered calcium concentration(P<0.001)and the expression levels of Wnt5a,Frizzled-2,and P-CaMK II in the IP area.In soy isoflavones-pretreated rats,calcium concentration,ROS and MDA levels,cell apoptosis rate,cerebral infarct volume,and expression levels of Wnt/Ca2+signaling pathway-related proteins were all significantly lower while SOD level was higher than those in rats in I/R model group.Conclusion Soy isoflavones can mitigate calcium overload in rats with cerebral I/R by inhibiting the Wnt/Ca2+signaling pathway.

Objective To investigate the changes and clinical significance of γδT cells and their major functional subsets γδT1 and γδT17 cells in the peripheral blood of patients with systemic lupus erythematosus (SLE). Methods Ratios and absolute numbers of γδT cells, γδT1 cells and γδT17 cells in peripheral blood were detected by flow cytometry in 21 SLE patients (SLE) group and 16 healthy controls (HC group). Correlations of γδT cells, γδT1 cells and γδT17 cells in the peripheral blood with clinical indicators were analyzed by Pearson correlation analysis. Results In SLE patients, the percentage of γδT cells to lymphocytes in the peripheral blood was (2.30±1.10)%, which was significantly lower than (3.30±1.51)% in the HC group (P < 0.05); the percentage of γδT1 cells to lymphocytes in peripheral blood was (0.93±0.67)%, which was significantly lower than (2.09±1.21)% in the HC group (P < 0.001); the ratio of γδT17 cells to lymphocytes in the peripheral blood was (1.53±2.82)‰, which showed an increasing trend when compared to (0.18±0.12)‰ in the HC group (P>0.05). The absolute number of γδT cells in the peripheral blood in SLE patients was (2.88±2.18)×10⁴/mL, which was significantly lower than (7.96±3.96)×10⁴/mL in the HC group (P < 0.000 1); the absolute number of γδT1 cells in the peripheral blood in SLE patients was (1.20±1.17)×10⁴/mL, which was significantly lower than (5.05±3.04)×10⁴/mL in the HC group (P < 0.000 1); the absolute number of γδT17 cells in the peripheral blood in SLE patients was (1.03±1.08)×10³/mL, which was significantly higher than (0.40±0.24)×10³/mL in the HC group (P < 0.05). The proportion of γδT17 cells in the peripheral blood of SLE patients was positively correlated with the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) (r=0.59, P < 0.01) and erythrocyte sedimentation rate (r=0.65, P < 0.01), while the absolute number of γδT17 cells was positively correlated with the SLEDAI score (r=0.59, P < 0.01) and negatively correlated with the complement C3 level (r=-0.53, P < 0.05). Conclusion The significant decrease in the number of γδT cells in the peripheral blood of SLE patients is mainly due to the reduction of its γδT1 cell subset; γδT17 cells play an important role in the pathogenesis and activity of SLE.

ObjectiveTo investigate the effect of Xinfeng capsules on immunoinflammatory indicators in patients with rheumatoid arthritis （RA） due to spleen deficiency and dampness exuberance. MethodA total of 102 patients were randomly divided into control group and observation group according to the random number table method， with 51 cases in each group. All patients were treated with methotrexate tablets， while those in the observation group received additional Xinfeng capsules. The course of treatment in both groups was 12 weeks. The 28-joint disease activity score （DAS28）， visual analogue scale （VAS） scores， morning stiffness time， erythrocyte sedimentation rate （ESR）， high-sensitivity C-reactive protein （hs-CRP）， rheumatoid factor （RF）， anti-cyclic citrullinated peptide （anti-CCP） antibody， vascular endothelial growth factor （VEGF）， and serum amyloid A （SAA） of the two groups before and after treatment were compared. The efficacy and incidence of adverse events were compared between the two groups. The Apriori association rule model and random walk model were constructed to evaluate the effect of Xinfeng capsules in improving hs-CRP， ESR， RF， SAA， VEGF， and anti-CCP. ResultThere were no dropouts in this study. There was no statistical difference in the indicators between the two groups before treatment. After 12 weeks of treatment， the total effective rate in the observation group was 90.19% （46/51）， which was higher than 74.51% （38/51） in the control group （χ²＝4.320，P<0.05）. DAS28， VAS score， and morning stiffness time in the observation group were improved compared with those in the control group （P<0.05）. Apriori association rule model results showed that the application of Xinfeng capsules in the observation group had a strong correlation with the reduction of RF， ESR， hs-CRP， SAA， and VEGF. The results of the random walk model showed that the improvement coefficients of hs-CRP， ESR， RF， SAA， and VEGF in the observation group were all better than those of the control group， and the improvement coefficient of anti-CCP in the control group was better than that of the observation group. The improvement degree of hs-CRP， ESR， RF， SAA， and VEGF in the observation group was superior to that of the control group （P<0.05）. The incidence of adverse reactions in the observation group was lower than in the control group （χ²＝4.057，P<0.05）. ConclusionOn the basis of the treatment with methotrexate tablets， Xinfeng capsules can effectively improve the immunoinflammatory level in RA due to spleen deficiency and dampness exuberance and reduce the incidence of adverse reactions.

Objective:To investigate the efficacy and safety of ixazomib-based therapy for multiple myeloma.Methods:The data of 32 patients with multiple myeloma treated with isazomib-based regimen in the Affiliated Hospital of Jining Medical University from December 2020 to December 2021 were retrospectively analyzed. Among 32 patients, 17 cases were relapsed/refractory, and the remaining 15 cases had initial treatment. The treatment regimens included ID (isazomib + dexamethasone), IRD (isazomib + lenalidomide + dexamethasone) and ICD (isazomib + cyclophosphamide + dexamethasone). The short-term curative effect and adverse reactions of relapsed/refractory patients and patients at initial onset were analyzed.Results:The overall response rate (ORR) of relapsed/refractory patients was 52.9% (9/17), of which 6 cases achieved complete remission (CR), 2 cases achieved very good partial remission (VGPR) and 1 case achieved partial remission (PR). The ORR of refractory patients receiving bortezomib therapy was 40.0% (4/10). The ORR of patients at initial onset who could be evaluated the curative effect was 100.0% (14/14), including 9 cases of CR, 2 cases of VGPR and 3 cases of PR. After treatment, 2 patients (6.2%) had grade Ⅲ-Ⅳ adverse events (1 case of herpes zoster and 1 case of thrombocytopenia), and none of the patients had grade Ⅲ-Ⅳ peripheral neuropathy.Conclusion:Isazomib is effective and safe in the treatment of initially treated and relapsed/refractory multiple myeloma.

Objective:To evaluate the safety and efficacy of daratumumab in the treatment of multiple myeloma (MM).Methods:The clinical data of 19 MM patients treated with daratumumab alone or in combination with chemotherapy regimens from June 2021 to December 2021 in the Affiliated Hospital of Jining Medical College were retrospectively analyzed, of which 2 patients received daratumumab alone, 6 cases received daratumumab combined with lenalidomide+dexamethasone (DRD) regimen, 1 case received daratumumab combined with liposomal doxorubicin+dexamethasone (DVD) regimen, 2 case received daratumumab combined with dexamethasone+cyclophosphamide+etoposide+cisplatin (DECP) regimen, 3 cases received daratumumab combined with isazomib+dexamethasone (ID) regimen, 2 cases received daratumumab combined with bortezomib+dexamethasone (BD) regimen, and 3 cases received daratumumab combined with dexamethasone (DD) regimen. The efficacy and incidence of adverse effects were analyzed.Results:Among the 19 patients, 8 had complete remission (CR), 1 had very good partial remission (VGPR), 5 had partial remission (PR), 1 had stable disease (SD), and 4 had progressive disease (PD). The overall response rate (ORR) was 73.7% (14/19). The median progression-free survival (PFS) time was 10.42 months (95% CI 8.04-12.79 months) and the median overall survival (OS) time was 52.06 months (95% CI 37.85-66.27 months). The main adverse reactions during treatment were grade 3 neutropenia in 3 cases, grade 3 lymphopenia in 3 cases, grade 2 anemia in 5 cases, grade 2 nausea and vomiting in 7 cases, and infusion-related adverse reactions in 7 cases. Conclusions:Daratumumab-based chemotherapy regimens for the treatment of MM patients can achieve great efficacy with good safety and tolerability.

Objective:To explore the clinical value of single-hole laparoscopic percutaneous extraperitoneal closure operation using a Kirschner wire assisted double-hook water-injection hernia needle in treating complicated pediatric oblique inguinal hernia.Methods:The clinical data of 366 children with oblique inguinal hernia treated in the Department of Urology Surgery, Children′s Hospital of Nanjing Medical University from December 2020 to October 2021 were retrospectively analyzed.According to the surgical methods, the children were divided into the ordinary crochet needle group and the Kirschner wire assisted group.Children treated by a single-port laparoscopic double hook water-injection hernia crochet needle (309 cases) were classified into the ordinary crochet needle group.Children treated by a single-port laparoscopic Kirschner wire assisted double hook water-injection hernia crochet needle (57 cases) were included in the Kirschner wire assisted group.The independent sample t-test and rank sum test was used to compare the relevant clinical indicators between the two groups. Results:Compared with the ordinary crochet needle group, children in the Kirschner wire assisted group were younger at surgery[(2.87±1.88) years vs.(4.91±2.39) years] and had larger hernia sacs [17 303.89(8 622.49, 37 295.42) mm 3vs.9 650.97(3 849.24, 17 539.51) mm 3]. The differences in the age at surgery and hernia sac volume were statistically significant ( t=-5.407, Z=-4.218; all P<0.001). There was no significant difference in body mass index between the 2 groups ( P>0.05). Taking hernias with sac volume >10 000 mm 3 as huge hernias, there were 70.18%(40/57 cases) and 47.25%(146/309 cases) of huge hernias in the Kirschner wire assisted group and the ordinary crochet needle group, respectively.The overall operation time of the Kirschner wire assisted group was significantly longer than that of the ordinary crochet needle group[(20(15, 20) min vs.15(15, 20) min] ( Z=-2.842, P<0.05). However, the operation time for huge oblique hernias with sac volume >10 000 mm 3 was not statistically significant between the 2 groups ( P>0.05). No recurrence in both groups was found during 6-16 months of follow-up. Conclusions:For complicated oblique inguinal hernia in children with a huge hernia or obvious retroperitoneal folds at the internal ring and heavy scar adhesion between the hernia sac and abdominal wall, the insertion of a Kirschner wire can help the hernia crochet needle to traverse the vas de-ferens and spermatic cord vessels smoothly.As a single port laparoscopic operation, the Kirschner wire assisted hernia crochet needle requires no addition of trocar holes and leaves only a small surgical scar.With good feasibility and safety, it is applicable for clinical popularization.

Porphyromonas gingivalis peptidylarginine deiminase (PPAD), an isoenzyme of animal endogenous peptidylarginine deaminase, is secreted by the Por system and catalyzes the citrullination of arginine. Recent studies have found that PPAD can affect the formation of Porphyromonas gingivalis biofilm and reduce the body′s immune defense function, which is related to the occurrence and development of many diseases such as periodontal diseases and rheumatoid arthritis. In this paper, we reviewed the molecular characteristics of PPAD, including the genetic and functional characteristics, as well as the mechanisms related to the inflammatory and autoimmune diseases. We also pointed some issues that should be pay attention to in the further study.

Objective:To investigate the clinical efficacy of autologous peripheral blood hematopoietic stem cell transplantation (HSCT) in treatment of lymphoma.Methods:The clinical data of 41 lymphoma patients undergoing autologous peripheral blood HSCT at the Affiliated Hospital of Jining Medical University between January 2014 to December 2020 were retrospectively analyzed. There were 6 cases of Hodgkin lymphoma and 35 cases of non-Hodgkin lymphoma. The mobilization regimens included chemotherapy drugs + granulocyte colony-stimulating factor (G-CSF) + thrombopoietin (TPO) or chemotherapy drugs + G-CSF. The pre-conditioning schemes before transplantation were listed as follows: BEAM (mustine + cytarabine + etoposide + melphalan) regimen + decitabine in 26 patients, BEAM regimen in 12 patients, BEAM regimen + chidamide in 3 patients. The progression-free survival (PFS), overall survival (OS), related complications, prognoses after transplantation were observed. The effects of clinical staging, B symptom,International Prognostic Score Index (IPI), extranodal involved sites, hemoglobin (Hb), lactic dehydrogenase (LDH), β 2-microglobulin (β 2-MG), transplantation regimen and the status before transplantation on PFS and OS after transplantation were evaluated. Results:Among 41 patients, 37 patients (90.24%) achieved complete remission (CR), 2 patients (4.88%) achieved partial remission (PR) and 2 patients loss assessment data (4.88%) before autologous peripheral blood HSCT. The median karyocyte count was 12.74×10 8 /kg [(3.91-22.68)×10 8/kg] in 24 patients with the complete data of stem cell collection, the median CD34 positive cell count was 6.74×10 6/kg [(0.91-50.47)×10 6/kg]. All 41 patients had hematologic reconstruction. The median time of platelet implantation was 11 d (7-32 d) and the median time of granulocyte implantation was 9 d (8-16 d). All patients achieved CR after transplantation and no one case had transplantation-related death. By the end of follow-up, 33 cases (80.49%) had no progression of disease, 8 cases (19.51%) died. The OS rates of 12-month, 24-month and 72-month were 93.4%, 85.3% and 60.9%, respectively after transplantation. The PFS rates of 12-month, 24-month and 72 month were 93.3%, 84.0% and 84.0%, respectively. Median PFS and OS had not been reached. There were no statistically significant differences in the PFS and OS of patients with different gender, clinical staging, B symptom, IPI score, extranodal involved sites, Hb, LDH, β 2-MG and the status before transplantation(all P > 0.05) . The PFS and OS of patients receiving BEAM regimen + decitabine were better than those of patients receiving BEAM regimen alone (all P < 0.05). Conclusions:Autologous peripheral blood HSCT is effective in treatment of lymphoma. Moreover, BEAM regimen + dicitabine preconditioning regimen can achieve longer survival time compared with BEAM regimen alone.

Objective: To explore the plasma level change of soluble tumor necrosis factor related apoptosis inducing ligand (sTRAIL) in patients with systemic lupus erythematosus (SLE) and its clinical significance. Methods: Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expressions of TRAIL and TRAIL receptors-1 (TRAIL-R1) and TRAIL-R2 in the peripheral blood mononuclear cells (PBMCs) derived from active SLE patients (n=26) and healthy controls. Enzyme linked immuno sorbent assay (ELISA) was used to detect the plasma level of sTRAIL in the active SLE patients (n=42), healthy controls (n=21). Pearson correlation analysis was used to analyze the correlation of sTRAIL with clinical and laboratory parameters. Results: The plasma levels of sTRAIL [(82±5) pg/ml] in SLE were significantly higher than that in healthy controls [(49±3) pg/ml], the difference was statistically significant (t=4.10, P<0.01). The plasma levels of sTRAIL in SLE with inactive disease [(92±14) pg/ml], mild active disease [(80±9) pg/ml], moderate active disease [(74±12) pg/ml] and severe active disease [(83±8) pg/ml] were higher than healthy controls, the difference was statistically significant (H=18.07, P<0.01). The mRNA levels of TRAIL and TRAIL-R2 in PBMCs derived from SLE patients were significantly higher than those in healthy controls [(1.04±0.08) vs (1.80±0.25), t=2.10, P<0.05 and (1.07±0.12) vs (2.08±0.21), t=3.27, P<0.01]. In SLE with moderate and severe active disease, plasma sTRAIL levels were associated with the 24 hours urine protein. Conclusion Plasma sTRAIL may be predictors of SLE disease activity and TRAIL pathway may be a new potential target of SLE treatment.