Osteoclasts were isolated mechanically from neonatal rat long bones and cultured on glass or devitalized human cortical bone slices. Both morphological and functional studies showed that the isolated cells were of typical osteoclasts, because; 1. they attached rapidly on the matrix and had pseudopodial activity when cultured on glass;2. they had multinuclei,most of them contained 3-10 nuclei;3. calci-tonin and clacitonin gene related peptide directly inhibited the cellular activity; 4. they showed high acid phosphatase activity; 5. resorption lacunae could be found when the cells cocultured with devitalized bone slices.

BACKGROUND: Recent data suggested that Notch signal pathway plays important regulatory effects in peripheral transplantation immunological response, promotes differentiation of regulatory T cells, induces antigen specific immune tolerance. We proposed that Notch/Notch ligand may play important roles in MHC/TCR interface.OBJECTIVE: To construct the eukaryotic expression vector of rat Deltai gene (Notch ligand), and to examine its expression in dendritic cells.METHODS: The complete encoding cDNA of rat-Delta1 was isolated from bone marrow cells by reverse transcription-polymerase chain reaction (RT-PCR) and this gene was recombined into pcDNA3.1(+) plasmid vector.pcDNA3.1/Delta1 plasmid was transfected into rat dendritic cells with lipofectamine gene transfection method.RESULTS AND CONCLUSION: Double enzyme digestion detection demonstrated that Delta 1 had been successfully constructed in Hindilll and xbal of pcDNA3.1. A positive clone pcDNA3.1/Delta1 was delivered to Shanghai Sangon Biological Engineering Technology & Services Co., Ltd. for sequencing. Sequencing results were identical to Delta1 gene sequence in Genebank, with correct reading frame. The Delta 1 gene-transfected dendritic cells showed similar morphology as their parent cells. Western blotting assay detected that Delta 1 expression was significantly increased in cells. The eukaryotic expression vector pcDNA3.1/Delta1 was constructed, and significant increase of Delta 1 expression was detected after transfection.

Objective To explore the significance of peritubular capillary C4d deposition in histopathological changes, renal function and prognosis of the patients with antibody-mediated chronic rejection (AMCR). Methods Deposition of C4d in the kidney was examined by irnmunohistochemistry on routine paraffin-embedded sections using anti-C4d polyclonal antibody. Seventy-seven patients were divided into C4d+ group (n = 35) and C4d- group (n = 42). The relationship of C4d and renal function,histopathological changes and prognoses of allografts were analyzed. Results The number of patients with tubular atrophy and glomerular basement membrane proliferation in C4d+ group was significantly more than that in C4d group (P＜0.05). Mean serum creatinine level was significantly higher in C4d+ group than in C4d- group 12 months after renal transplantation [(379.1 + 260.2)μmol/L vs (260.5 + 175.3) μmol/L, P＜0.05]. According to Kaplan-Meier analysis, the one-year graft survival rate was lower in the C4d+ group (62.9% ) than in the C4d- group (83.3% ) (logrank P＜0.05). Conclusion Patients with C4d deposition are associated with tubular atrophy and glomerular basement membrane proliferation. The serum creatinine level in C4d+ patients was significantly higher than in C4d- group at the 12th month after transplantation. More patients with C4d deposition lost their grafts during the study period.

ObjectiveTo investigate the efficacy of rapamycin combined with CsA/Tacrolimus (Tac) in chronic allograft nephropathy (CAN).MethodsFifty-three cases of CAN accepted the quadruple immunosuppressive drug program,which contained rapamycin combined with CsA/Tac and MMF and prednisone,and CsA/Tac and MMF were reduced to the original amount of 25％ to 50％.After treatment for 12 months,more relevant indicators,including serum creatinine,glomerular filtration rate,serum cholesterol,triglycerides,urinary protein,GPT and bilirubin and other changes were observed.ResultsIn the patients receiving quadruple regimen of rapamycin during 12 months,the blood Ccr was decreased from (161.51 ± 106.48)μmol/L before treatment to (126.51 ± 56.2)μmol/L after treatment for 6 months (P＜0.05) and to (123.43 ± 54.18)μmol/L after for 12 months (P＜0.01).The GFR was increased from (0.754 ± 0.302) ml/s before treatment to (0.952 ± 0.347)ml/s after treatment for 6 months (P＜0.05) and to (1.007 ± 0.394) ml/s after treatment for 12 months (P＜0.01).Cholesterol and triglycerides in patients had no significant change before and after treatment.The positive rate of proteinuria after treatment showed an increasing trend from 9.4％ before treatment to 26.4％ after treatment for 12 months.ConclusionThe quadruple program of rapamycin combined with CsA/FK506 and MMF can significantly improve Ccr and GFR in patients with CAN,but it can increase the incidence of proteinuria in patients:

PURPOSE: The definition of nodal pathologic complete response (pCR) after a neoadjuvant chemotherapy (NAC) just included the evaluation of axillary lymph node (ALN) without internal mammary lymph node. This study aimed to evaluate the feasibility of internal mammary-sentinel lymph node biopsy (IM-SLNB) in patients with breast cancer who underwent NAC. METHODS: From November 2011 to 2017, 179 patients with primary breast cancer who underwent operation after NAC were included in this study. All patients received radiotracer injection with modified injection technology. IM-SLNB would be performed on patients with internal mammary sentinel lymph node (IMSLN) visualization. RESULTS: Among the 158 patients with cN+ disease, the rate of nodal pCR was 36.1% (57/158). Among the 179 patients, the visualization rate of IMSLN was 31.8% (57/179) and was 12.3% (7/57) and 87.7% (50/57) among those with cN0 and cN+ disease, respectively. Furthermore, the detection rate of IMSLN was 31.3% (56/179). The success rate of IM-SLNB was 98.2% (56/57). The IMSLN metastasis rate was 7.1% (4/56), and all of them were accompanied by ALN metastasis. The number of positive ALNs in patients with IMSLN metastasis was 3, 6, 8, and 9. The pathology nodal stage had been changed from pN1/pN2 to pN3b. The pathology stage had been changed from IIA/IIIA to IIIC. CONCLUSION: Patients with visualization of IMSLN should perform IM-SLNB after NAC, especially for patients with cN+ disease, in order to complete lymph nodal staging. IM-SLNB could further improve the definition of nodal pCR and guide the internal mammary node irradiation.
Biopsy ; Breast Neoplasms* ; Breast* ; Drug Therapy* ; Humans ; Lymph Nodes ; Neoadjuvant Therapy ; Neoplasm Metastasis ; Pathology ; Polymerase Chain Reaction ; Sentinel Lymph Node Biopsy*

Objective To analyze the clinical diagnosis and treatment data of 20 children with hypophosphatemic rickets (HR) in order to improve the clinical diagnosis and treatment of HR.Methods The retrospective analysis of clinical data of 20 cases with HR who were hospitalized at Children's Hospital of Nanjing Medical University from May,2010 to April,2016 was performed to summarize the clinical characteristics.All patients were analyzed for the phosphate regulating gene with homologies to endopeptidase on the X chromosome(PHEX) gene by direct sequencing.If no mutations were detected,multiplex ligation-dependent probe amplification analysis was performed.Results All of the 20 cases with HR showed different degrees of growth retardation and typical X-ray rickets.After treatment,the clinical features were improved.Height standard deviation score (HSDS) was improved significantly with longer treatment time,and the difference was statistically significant(P =0.027).There was a correlation between the blood phosphorus fluctuation and secondary hyperparathyroidism(P ＜ 0.05).Nineteen cases had PHEX gene mutations.Truncating mutations was the most frequent mutation type,and 4 new mutations were found.Conclusions Clinical characteristics,laboratory test results and X-ray examination are important clinical index for the diagnosis of HR,and PHEX gene test can be used as an important auxiliary diagnostic tool.Early diagnosis and treatment can significantly improve the clinical manifestations of the patients.

Objective To evaluate the effects of two-step approach combined with unprotected perineum method on the incidence uf neonatal swallowing syndrome.Methods A total of 360 patients were randomly divided into three groups:A,B and C,with 120 cases in each group.Group A was provided traditional midwifery approach:midwifery staff manually controlled the delivery rate of fetal head with hands,assisted to deliver fetal shoulders after fetal head was delivered,and quickly used hands to clean up the completely delivered newborn and routinely sucked mucus from mouth and nose using aurilave.Group B and C were provided two-step approach combined with unprotected perineum method:midwifery staff did not interfere delivery with hands,paused after the fetal head was delivered,delivered the fetal shoulders until next contraction.Group B was cleaned up respiratory tract like group A,group C was only provided surface cleaning or squeezing nose and mouth using hands.The incidence of neonatal swallowing syndrome and the number of vomiting were recorded within 24 hours after birth among three groups,and the differences among three groups were compared.Results The incidence of neonatal swallowing syndrome in group A was higher than those in groups B(x2=6.504,P=0.011)and C(x2=7.533,P=0.006);there was no significant difference between group A and B in neonatal swallowing syndrome(x2=0.042,P=0.837).There were significant differences in vomiting within 24 hours among three groups(F=10.185,P=0.006).There were significant differences in vomiting within 24 hours between group A and group B,and between group A and group C(P＜0.05);there was no significant difference between group B and group C(P＞0.05).Conclusion Inplementing the two-step approach combined with unprotected perineum method,the newborns can accomplish cleaning up respiratory tract naturally during natural delivery of fetal shoulders.

Objective:To investigate the relationship between peripheral blood helper T cell 1/helper T cell 2 (Th1/Th2), helper T cell 17/regulatory T cell (Th17/Treg) and efficacy of programmed death receptor 1 (PD-1) inhibitor in patients with advanced non-small cell lung cancer (NSCLC) combined with chronic obstructive pulmonary disease (COPD).Methods:The clinical data of 107 patients with advanced NSCLC combined with COPD who were admitted to Suzhou Ninth Hospital Affiliated to Soochow University from April 2021 to September 2022 were retrospectively analyzed. The patients were treated with PD-1 inhibitor, and they were categorized into the disease control group (82 cases) and the disease progression group (25 cases) according to the clinical efficacy. Th1/Th2, Th17/Treg and clinical data of patients before treatment were compared between the two groups, and logistic regression was used to analyze the independent influencing factors of patients' PD-1 inhibitor efficacy.Results:Th1/Th2 and Th17/Treg before treatment in the disease control group were higher than those in the disease progression group (12.49±1.14 vs. 7.04±1.06, t = 21.26, P < 0.001; 0.14±0.03 vs. 0.09±0.04, t = 6.72, P < 0.001). The proportions of patients with TNM stage Ⅳ, lymph node metastasis and brain metastasis in the disease progression group were higher than those in the disease control group (all P < 0.01). The results of multivariate logistic regression analysis showed that pre-treatment Th1/Th2 ( OR = 0.744, 95% CI 0.685-0.799, P < 0.001), pre-treatment Th17/Treg ( OR = 0.514, 95% CI 0.465-0.552, P < 0.001), TNM stage ( OR = 1.258, 95% CI 1.049-1.656, P = 0.048), lymph node metastasis ( OR = 1.790, 95% CI 1.223-2.734, P = 0.005), and brain metastasis ( OR = 1.640, 95% CI 1.184-2.348, P = 0.005) were independent influencing factors of PD-1 inhibitor efficacy in patients with advanced NSCLC combined with COPD. Conclusions:Patients with advanced NSCLC combined with COPD who have high Th1/Th2 and Th17/Treg before treatment have good outcomes with PD-1 inhibitor therapy.

Objective:To explore the serum levels of inflammatory cytokines and prognosis in severe acute infection children with glucose-6-phosphate dehydrogenase(G6PD) deficiency.Methods:A total number of 160 children with severe acute infections admitted to PICU of Guangxi Zhuang Autonomous Region Maternal and Child Health Hospital from June 2014 to December 2017 were selected as subjects in this study, including 80 children with G6PD deficiency(observation group) and 80 children without G6PD deficiency(control group). The changes of TNF-α, IL-6, IL-10 and CRP were dynamically monitored at 0-hour, 12-hour and 24-hour after admision, and the occurrences of sepsis, multiple organ dysfunction syndrome(MODS) were prospectively analyzed.Results:The levels of serum cytokines and CRP in the observation group were significantly higher than those in the control group at admission[TNF-α: (65.57±19.09) pg/ml vs.(46.53±20.34) pg/ml; IL-6: (98.90±29.02) pg/ml vs.(89.89±25.54) pg/ml; IL-10: (87.66±21.84) pg/ml vs.(76.34±19.01) pg/ml; CRP: (60.18±22.24) mg/L vs.(41.43±19.51) mg/L, respectively], and the differences between two groups were statistically significant( P<0.05). The levels of cytokines and CRP in the observation group were higher than those in the control group at 12 h and 24 h after treatment( P<0.01). Compared with the control group, the incidences of sepsis(82.50% vs 67.50%) and MODS(73.75% vs 58.75%) in the observation group increased, and the recovery rate(81.25% vs 92.50%) decreased, with statistical significance between two groups( P<0.05). Conclusion:Children with G6PD deficiency need to be paid more attention to inflammation, sepsis, MODS and the difficulty of treatment when they are infected.The potential mechanism may be related to oxidative stress.

Objective:To investigate the short-term efficacy of laparoscopic surgery after short-course radiotherapy followed by sequential chemotherapy combined with anti-programmed death-1 (PD-1) antibody therapy for locally advanced rectal cancer.Methods:The prospective study was conducted. The clinicopathological data of 30 locally advanced rectal cancer patients who were admitted to the Union Hospital of Tongji Medical College of Huazhong University of Science and Technology from November 2019 to September 2020 were selected. Patients underwent laparos-copic surgery after short-course radiotherapy followed by sequential chemotherapy combined with anti-PD-1 antibody therapy. Observation indicators: (1) situations of the enrolled patients; (2) situations of short-course radiotherapy followed by sequential chemotherapy combined with anti-PD-1 antibody therapy and adverse events; (3) preoperative evaluation and surgical situations; (4) postoperative situations and pathological examinations; (5) postoperative adjuvant chemo-therapy and follow-up. Follow-up was conducted using outpatient examination and telephone interview up to March 2022. Patients were followed up once every 3 weeks during the period of short-course radiotherapy followed by sequential chemotherapy combined with anti-PD-1 antibody therapy to detect the adverse events and patients were followed up once every 3 months during the first postoperative 2 years and once every 6 months thereafter to detect tumor recurrence and survival of patients. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers or percentages. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. Results:(1) Situations of the enrolled patients. A total of 30 patients were selected for eligibility. There were 17 males and 13 females, aged (57±16)years. Cases with preoperative primary tumor in stage cT3 and cT4 were 22 and 8, respectively. Cases with preoperative clinical lymph node metastasis in stage cN0, cN1, cN2 were 4, 16, 10, respectively. Cases in preoperative clinical stage Ⅱ and Ⅲ were 4 and 26, respectively. Of the 30 patients, there were 21 cases with positive circumferential margin and 12 cases with vascular invasion in extramural of rectum in the preoperative imaging evaluation. Distance from the distal margin of tumor to anal margin and tumor diameter of the 30 patients were 4.7(range, 1.9?9.0)cm and 5.4(range, 2.1?10.0)cm, respectively. There were 28 cases with mismatch repair proficient and 1 case with mismatch repair deficiency in tumor tissues. There was 1 case missing the data of mismatch repair in tumor tissues as failed in biopsy of pathological examination before the treatment. (2) Situations of short-course radiotherapy followed by sequential chemotherapy combined with anti-PD-1 antibody therapy and adverse events. All the 30 patients completed preoperative short-course radiotherapy successfully. Of the 30 patients, there were 3 cases not undergoing the sequential chemotherapy combined with anti-PD-1 antibody therapy and there were 24 cases undergoing 2 courses of the sequential chemotherapy combined with anti-PD-1 antibody therapy and 3 cases undergoing 1 course of the sequential chemotherapy combined with anti-PD-1 antibody therapy. The time interval between ending of radiotherapy and starting of chemotherapy combined with anti-PD-1 antibody therapy of the 27 patients was 12(range, 4?18) days. Cases with leukopenia, cases with endothelial hyperplasia of skin capillaries, cases with radiation proctitis, cases with anemia, cases with peripheral neurotoxicity, cases with neutropenia, cases with thrombocytopenia, cases with fatigue, cases with anorexia, cases with abnormal liver function, cases with hypothyroidism were 24, 22, 21,20, 18, 16, 16, 13, 10, 9, 2 in the 30 patients during the preoperative short-course radiotherapy followed by sequential chemotherapy combined with anti-PD-1 antibody therapy. Cases with the above adverse events were improved after symptomatic treatment. (3) Preoperative evaluation and surgical situations. Seven of the 30 patients were in clinical complete remission after preoperative multidisciplinary evaluation and the other 23 patients were not in clinical complete remission. Twenty-seven of the 30 patients underwent laparoscopic radical resection of rectal cancer and 3 patients not undergoing the sequential chemotherapy combined with anti-PD-1 antibody therapy did not undergo surgery. The time interval between ending of chemotherapy combined with anti-PD-1 antibody therapy and the surgery of the 27 patients were 14(range, 5?141)days. Of the 27 cases, there were 13 cases and 14 cases with 0 and 1 of the preoperative Eastern Cooperative Oncology Group score, respectively, and there were 24 cases undergoing low anterior proctectomy and 3 cases undergoing abdominoperineal excision. The operation time and volume of intra-operative blood loss of the 27 cases were (182±36)minutes and 30(range, 10?150)mL, respectively. Of the 27 cases, there were 16 cases with protective ileostomy and 24 cases with anal preservation. (4) Postoperative situations and pathological examinations. The time to postoperative first flatus, time to postoperative initial liquid food intake and duration of postoperative hospital stay of the 27 patients undergoing surgery were 2(range, 1?4)days, 3(range, 2?5)days and 8(range, 7?16)days, respectively. Five of the 27 patients had postoperative grade Ⅰ?Ⅱ complications, including 2 cases with incision infection, 1 case with abdominal infection, 1 case with incision hemorrhage and 1 case with venous thrombosis in left lower limb intermuscular. Cases with postoperative complica-tions were improved after symptomatic treatment. Results of postoperative pathological examina-tion showed that the rate of pathologic complete response in 27 patients was 48.1%(13/27). Of the 27 cases, cases in grade 0, grade 1, grade 2, grade 3 of the tumor regression grading were 13, 5, 7, 2, respectively, cases in stage T0, stage Tis, stage T2, stage T3 of the tumor T staging were 13, 1, 5, 8, respectively, cases in stage N0, stage N1, stage N2 of the tumor N staging were 19, 6, 2, respectively, cases in stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲ of the tumor TNM staging were 14, 0, 5, 8, respectively. The number of lymph node dissected of the 27 patients was 15(range, 3?29). Of the 27 patients, there was 1 case with positive circumferential margin and 26 cases achieving R 0 resection. None of the 27 patients underwent secondary operation or perioperative death. (5) Postoperative adjuvant chemotherapy and follow-up. Of the 27 patients undergoing surgery, 21 cases underwent post-operative adjuvant chemotherapy, with the cycles of 4(range, 1?6). All the 27 patients were followed up for 20(range, 20?29)months. During the follow-up, 3 cases not achieving pathological complete response had tumor recurrence and no patient died. The disease free survival rate of the 27 patients was 88.9%. Conclusion:Laparoscopic surgery after short-course radiotherapy followed by sequential chemotherapy combined with immunotherapy for locally advanced rectal cancer is safe and feasible, with satisfied short-term efficacy.