Objective To construct a prokaryotic expression vector of of the long QT syndrome(LQTS)associated C-terminal lobe of calmodulin(CaM)mutant E141G(C-lobeE141G)and to identify the expression,purification,and activity of C-lobeE141G.Methods A cDNA fragment was inserted into a PGEX-6p-3 plasmid vector and transferred into Escherichia coli BL21 receptor cells,and glutathione-S-trans-ferase(GST)fusion protein was induced by isopropyl thio-β-D galactoside(IPTG).Glutathione-Sepharose 4B beads were used to separate and purify GST-C-lobeE141G.After removing the GST label with protease,the purity and concentration of purified C-lobeE141G were detected using SDS-PAGE and BCA,respectively.The activity of purified C-lobeE141G was detected using the GST pull-down method and patch clamp technique.Results GST-C-lobeE141G fusion protein was highly expressed,and C-lobeE141G with high purity and concentration was obtained.The purified C-lobeE141G protein not only bound to CaV1.2 calcium channels,but also rescued the channel activity from run-down in the ventricular myocytes of rat hearts.Conclusion This study successfully constructed a prokaryotic expression vector of C-lobeE141G,which provides a material basis for the study of the mechanism of LQTS mediated by C-lobe mutations in CaM.

Objective To investigate the inhibitory effect of GsMTx4 on pain sensation in rats during tooth movement,open field ex-periments and immunohistochemical staining by conducting.Methods Fifty-eight male SD rats aged 5-6 weeks were selected and a right maxillary tooth movement model was constructed.The rats were randomly divided into two groups:group A(control group)and group B(GsMTx4 group).They were also divided into 5 subgroups according to 4 h,1 d,3 d,5 d,and 7 d.The left maxillary of the rats in group A was used as group C(blank control group).The open-field test was used to evaluate the pain level of rats.Immunohisto-chemical staining was used to observe the expression of nerve growth factor(NGF)and calcitonin gene-related peptide(CGRP)in peri-odontal tissues.Results At 4 h,1 d,and 3 d,the total distance traveled by rats in group B within 5 minutes in the open-field was higher than that of group A(P＜0.05).At 4 h and 1 d,the percentage of time in the center of rats in group B was higher than that in group A(P＜0.05).The expressions of NGF and CGRP in groups A and B were higher than those in group C on 1 d,3 d,5 d,and 7 d(P＜0.05).The expression of NGF and CGRP in group B were lower than those in group A at 4 h,1 d,and 3 d(P＜0.05).Conclu-sion GsMTx4 can inhibit the expression of NGF and CGRP during tooth movement and relieve the pain caused by tooth movement.

Objective:To evaluate the efficacy and safety of CD30 antibody-drug conjugates (ADC) brentuximab vedotin (BV) combined with chemotherapy in children with refractory or relapsed classic Hodgkin′s lymphoma (R/R cHL).Methods:Clinical data (including age, gender, B symptoms, clinical stage, previous treatment, etc.) of the 10 R/R cHL children diagnosed and treated at Beijing Children′s Hospital Affiliated to Capital Medical University from October 2021 to August 2023 were analyzed retrospectively. According to the different intensity of chemotherapy drugs, the dose of BV applied in the same course of treatment was 1.8 mg/kg for BV applied once every 3 weeks, and 1.2 mg/kg for BV applied once every 2 weeks. All 10 patients received at least 2 cycles of BV combined with chemotherapy and were evaluated every 2 cycles. The patients were followed up until May 31, 2024. The infusion reactions and adverse reactions after treatment were recorded.Results:In all 10 patients, there were 7 males and 3 females, the age ranged from 5.3-16.9 years, and there were 6 cases of refractory and 4 cases of relapsed. There were 6 cases of nodular sclerosis type, 2 cases of mixed cell type, 1 case of lymphocyte-rich type, and 1 case of lymphodepletion type. There were 5 cases of stage Ⅳ and 5 cases of stage Ⅲ. Previous treatment was mainly chemotherapy, 4 cases received radiotherapy and 1 case received programmed cell death protein 1 (PD-1) antibody therapy. The follow-up time ranged from 9 to 27 months. A total of 43 courses with 49 doses of BV alone or combined with chemotherapy were recorded, and the number of courses was 2 to 10 times. All 10 children responded to the treatment, and 9 achieved complete remission. BV infusion was successfully completed in all cases. A total of 28 cases of grade 3 or above adverse events were recorded, mainly myelosuppression, all of which were related to chemotherapy and did not affect sequential treatment.Conclusion:Brentuximab vedotin has demonstrated efficacy and a tolerable safety profile in the treatment of refractory and relapsed CD30-positive Hodgkin′s lymphoma in children.

Objective To investigate the effect of the Piezo1 channel on tension-side angiogenesis and osteogenic remodeling during orthodontic tooth movement,so as to provide an experimental basis for accelerating orthodontic peri-odontal tissue remodeling.Methods This study was approved by the Animal Ethics Committee.Sixty healthy male Sprague-Dawley rats with bilateral maxillary incisors as the anchorage were selected,and nickel titanium tension springs were used to apply 0.5 N of force to the right maxillary first molar of the rats and construct an orthodontic tooth movement model.The rats were randomly divided into three groups(n=20 per group).On Days 0 and 8 of force appli-cation,equal volumes of saline(control group),100 μmol/L Piezo1 channel agonist(Yoda1 group),and 48 μmol/L Piezo1 channel inhibitor(GsMTx4 group)were injected into the buccal and palatal submucosa of the right maxillary first molar.Tooth movement distances were recorded on Days 1,3,7,and 14.Five rats from each group were sacrificed at each time point to obtain maxillary tissue samples.Hematoxylin and eosin(HE)staining was performed to observe the histophysiological changes in the tension-side periodontal tissues.Immunohistochemical staining was used to mark and count CD31-positive cells(microvascular quantification)and to detect the expression of osteocalcin(OCN)in the tension side.Results Measurements of tooth movement distance showed that the Yoda1 group exhibited significantly in-creased tooth movement distances on Days 3,7,and 14(0.238±0.008 mm,0.406±0.011 mm,and 0.746±0.013 mm,respectively)compared to the control group(P＜0.05).In contrast,the GsMTx4 group showed significantly reduced tooth movement distances on Day 7(0.282±0.011 mm)and Day 14(0.578±0.008 mm)compared to the control group(P＜0.05).HE staining results indicated that the periodontal ligament space on the tension side gradually widened with the duration of force application and then gradually returned to normal,with visible osteoblasts.Quantitative analysis of CD31-positive cells(microvascular quantification)showed that the Yoda1 group had significantly increased numbers of blood vessels on Day 3(8.027±0.225)and Day 7(14.320±0.471)compared to the control group(P＜0.05),peaking on Day 7 and then gradually decreasing.The GsMTx4 group showed significantly fewer blood vessels in the periodontal ligament on the tension side on Day 3(6.013±0.177),Day 7(9.187±0.678),and Day 14(12.613±0.334)compared to the control group(P＜0.05).Immunohistochemical results indicated that the Yoda1 group had significantly increased OCN expression on Days 1,3,7,and 14 compared to the control group(P＜0.05),while the GsMTx4 group showed re-duced OCN expression on Days 7 and 14(P＜0.05).Conclusion Activation of the Piezo1 channel promotes orthodon-tic tooth movement,tension-side angiogenesis,and osteogenic remodeling,while inhibition of the Piezo1 channel produces the opposite effect.

Objective To study the brain functional connectivity(FC)changes in patients with normal tension glau-coma(NTG)and healthy volunteers using FC technique of resting-state functional magnetic resonance imaging(rs-fMRI)based on V1 region seed point(ROI),so as to explore the pathogenesis and early diagnosis of NTG.Methods Fourteen NTG patients(NTG group)who met the inclusion criteria and 14 healthy controls(HCs group)were enrolled.The clinical data of all subjects were collected,and rs-fMRI was performed in both groups.The magnetic resonance data was pre-pro-cessed by software,and bilateral A1 regions were taken as the ROI to analyze their correction with the whole brain voxel time series and obtain the FC value between the ROI and the whole brain by comparison of FC values in resting state be-tween the groups.Pearson correlation analysis was used to explore the relationship between FC value in the brain regions with significant differences with the ROI and clinical variables in the NTG group.Results Compared with the subjects in the HCs group,there were no statistically significant differences in age,gender,body weight,cup-disc ratio and 24 h mean intraocular pressure of patients in the NTG group(all P＞0.05),and there were statistically significant differences in the best corrected visual acuity(BCVA)of both eyes and peripapillary retinal nerve fiber layer thickness(RNFLT)(all P＜0.05).The Pearson correlation analysis showed that FC value of the brain regions with abnormal FC to V1 region were cor-related with RNFLT in the NTG group(P＜0.05).ROI1-left superior frontal gyrus,ROI1-right superior frontal gyrus,ROI2-left cingulate gyrus and ROI2-right middle frontal gyrus were significantly positively correlated with RNFLT(all P＜0.05).Compared with the HCs group,the brain regions with reduced FC to the right ROI in the NTG group were the left superior frontal gyrus and right superior frontal gyrus;the brain regions with reduced FC to the left ROI were the left cingulate gyrus and right middle frontal gyrus.Conclusion Compared to healthy individuals,NTG patients have significant changes in the functional connections between certain specific brain regions and V1 region,including bilateral superior frontal gyrus,left cingulate gyrus,and middle frontal gyrus.The changes in brain functional activity may be caused by visual dysfunction caused by NTG,leading to functional impairment of the visual and cognitive emotion processing brain regions,which may be one of the potential neuropathological mechanisms in NTG patients.

Objective:To investigate the clinical efficacy of redo rectal resection and coloanal anastomosis.Methods:The retrospective and descriptive study was conducted. The clinicopatholo-gical data of 49 patients who underwent redo rectal resection and coloanal anastomosis for the treatment of local recurrence of tumors and failure of colorectal or coloanal anastomosis after rectal resection in the Sixth Affiliated Hospital of Sun Yat-sen University from November 2012 to December 2021 were collected. There were 32 males and 17 females, aged 57(range,31-87)years. Redo rectal resection and coloanal anastomosis was performed according to the patient′s situations. Observa-tion indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distri-bution were represented as M( Q1, Q3) or M(range). Count data were described as absolute numbers or percentages. Results:(1) Surgical situations. All 49 patients underwent redo rectal resection and coloanal anastomosis successfully, with the interval between the initial surgery and the reopera-tion as 14.2(7.1,24.3)months. The operation time and volume of intraoperative bold loss of 49 patients in the redo rectal resection and coloanal anastomosis was 313(251,398)minutes and 125(50,400)mL, respectively. Of the 49 patients, there were 38 cases receiving laparoscopic surgery including 12 cases with transanoscopic laparoscopic assisted surgery, 11 cases receiving open surgery including 2 cases as conversion to open surgery, there were 20 cases undergoing Bacon surgery, 14 cases undergoing Dixon surgery, 12 cases undergoing Parks surgery, 2 cases undergoing intersphincter resection and 1 case undergoing Kraske surgery, there were 20 cases undergoing rectum dragging out excision and secondary colonic anastomosis, 13 cases undergoing dragging out excision single anastomosis, 12 cases undergoing rectum dragging out excision double anastomosis, 4 cases undergoing first-stage manual anastomosis, there were 21 cases with enterostomy before surgery, 16 cases with prophylactic enterostomy after surgery, 12 cases without prophylactic enterostomy after surgery. The duration of postoperative hospital stay of 49 patients was (14±7)days. (2) Postoperative situations. Fifteen of 49 patients underwent postoperative complications, including 8 cases with grade Ⅱ Clevien-Dindo complications and 7 cases with ≥grade Ⅲ Clevien-Dindo complications. None of 49 patient underwent postoperative transferring to intensive care unit and no patient died during hospitalization. Results of postoperative histopathological examination in 23 patients with tumor local recurrence showed negative incision margin of the surgical specimen. (3) Follow-up. All 49 patients underwent post-operative follow-up of 90 days. There were 42 cases undergoing redo rectal resection and coloanal anastomosis successfully and 7 cases failed. Of the 37 patients with enterostomy, 20 cases failed in closing fistula, and 17 cases succeed. There were 46 patients receiving follow-up with the median time as 16.1(7.5,34.6)months. The questionnaire response rate for low anterior resection syndrome (LARS) score was 48.3%(14/29). Of the patients who underwent redo coloanal anastomosis and closure of stoma successfully, there were 9 cases with mild-to-moderate LARS.Conclusion:Redo rectal resection and coloanal anastomosis is safe and feasible for patients undergoing local recurr-ence of tumors and failure of colorectal or coloanal anastomosis after rectal resection, which can successfully restore intestinal continuity in patients and avoid permanent enterostomy.

Objective:To investigate the feasibility of mandibular angle osteotomy combined with high-density polyethylene (MEDPOR) prosthesis implantation for one-time lower facial contouring of the protuberance and related surgical precautions.Methods:A total of 18 patients who were treated in the oral and maxillofacial surgery of Yangzhou First People＇s Hospital from June 2010 to August 2016 were selected to complete the mandibular angle osteotomy combined with MEDPOR prosthesis implantation in the lower part of the prosthetic plane. The patients were followed up regularly and the effect of surgery was observed.Results:All 18 patients were operated successfully with an average time of 2 hours. The effect of operation was satisfactory. Except for one patient whose hematoma caused local bruise to subside completely in 2 weeks, all the others were discharged smoothly in one week after operation. After 2 months to 3 years of follow-up, the effect was satisfactory.Conclusions:Mandibular angle osteotomy combined with MEDPOR prosthesis is safe and effective.

Objective To explore the the expression of Klotho mRNA and protein in placenta of macrosomia and its relationship with the birth weight of neonates. Methods The cases were from November 2014 to March 2015 in Shengjing Hospital of China Medical University, divided into 4 groups:the gestational diabetes with macrosomia group (GM), the gestational diabetes with normal birth weight group (GN), the normal pregnancy with macrosomia group (NM) and the normal pregnancy with normal birth weight group (NN). Klotho mRNA and protein expression in the placenta were detected by immunohistochemistry SP method, real-time fluorescent quantitative PCR and western blot, respectively, and were compared among the 4 groups. Results (1) Immunohistochemical detection showed the positive rate of Klotho protein was significantly higher in the placenta of GM (93%,28/30) than in the GN (73%,22/30;P<0.05). The positive rate was significantly higher in the placenta of NM (97%,29/30) than in the NN (80%,24/30;P<0.05). (2) Real-time fluorescent quantitative PCR showed the Klotho mRNA expression was significantly higher in the placenta of GM (4.3 ± 3.1) than in the GN (2.1 ± 2.4;P<0.05). The Klotho mRNA expression was also significantly higher in the placenta of NM (4.8 ± 3.4) than in the NN (2.6 ± 3.3;P<0.05). (3) Western blot showed the Klotho protein expression was significantly higher in the placenta of GM (1.27±0.90) than in the GN (0.64±0.24;P<0.05). It was also significantly higher in the placenta of NM (2.51±3.52) than in the NN (0.77±0.37;P<0.05). (4) There were no significant differences in the expression of Klotho mRNA and protein between GM and NM, GN and NN (P>0.05). Conclusions The up-regulation of Klotho gene may be associated with macrosomia. The relationship is not affected by the complication of gestational diabetes.

Objective To evaluate the relationship between placental expression of Gab1 and neonatal birth weight in mothers with gestational diabetes mellitus (GDM).Methods From the singleton and full-term cesarean delivered women in Shengjing Hospital Affiliated to China Medical University between October 2014 and May 2015,30 macrosomia babies with maternal GDM were selected as GDM macrosomia group,30 cases of GDM with normal neonatal birth weight as GDM normal group,30 cases without GDM but with macrosomia as normal macrosomia group,and 30 cases without GDM and with normal neonatal birth weight as the normal control group.Gab1 protein and mRNA expression in placentas were detected using immunohistochemistry,Western blot and real-time quantitative-polymerase chain reaction.Analysis of variance,LSD,Dunnett's T3,Chi-square test and Pearson's correlation analysis were used for statistical analysis.Results (1) Gab 1 protein location and positive expression rate:Gab 1 protein expression in human placenta tissue was located in the nucleus.The positive epression rate of Gab 1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group [93％(28/30),73％(22/30) vs 73％(22/30)] and those in the normal macrosomia group and GDM normal group were higher than in the normal control group[47％(14/30)](x2=4.320,4.320,4.444 and 4.444,all P＜0.05).(2) The expression levels of Gabl protein and mRNA:The expression level of Gab1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group (1.43 ± 0.58 vs 1.05 ± 0.67 and 0.95± 0.59),and that in the normal macrosomia group and GDM normal group were higher than in the normal control group (0.64±0.38) (LSD test,all P＜0.05).The expression levels of Gab1 mRNA showed the same trend as the expression levels of Gab1 protein in the four groups.(3) Gab 1 protein expression level was positively associated with neonatal birth weight (r=0.320,P=0.320).Conclusions The expression of Gab1 in placenta is involved in the regulation of birth weight in GDM mothers.

Objective:To investigate the effect of butyrate produced by bacterial metabolism on immune features of murine bone marrow-derived dendritic cells(BMDCs) and its potential mechanisms.Methods: BMDCs were prepared from bone marrow cells of C57BL/6 mice by being cultured with GM-CSF and IL-4.The expression of CD80,CD86,B7-DC and MHCⅡ on BMDCs and its pri-ming ability on the proliferation of OVA257-264 antigen specific CD8+T cell were analyzed by using Flow cytometry.The mRNA levels of IL-6 and IL-12 in BMDCs were detected by real-time fluorescence quantitative PCR(q-PCR).Simultaneously,Griess reaction and Western blot was used for analyzing the levels of NO2-in BMDCs culture supernatant and the ERK phosphortylation in BMDCs respectivly.Results:Butyrate could decrease the levels of CD80,CD86,MHCⅡand B7-DC,and downregulate the capability of BMDCs in priming the proliferation of CD8+T cells.Furthermore,the secretions of IL-6,IL-12,NO2-and the phosphorylation of ERK were sup-pressed.Conclusion:Butyrate down-regulats the immune functions of BMDCs via inhibition of ERK phosphorylation in TLR 4 signaling pathway.