Objective To investigate the effects of KangAi injection on apoptosis and proliferation of rectal cancer Lovo cells in vitro.Methods SubG1 and Annexin -V /PI were used to examine the apoptosis of Lovo cells in vitro.Real -time PCR was used to examine the expression of Caspase -3 and Bcl -2 at mRNA level.Proliferation protein ki -67 was detected by flow cytometry.Results Lovo cells treated with KangAi injection 180μL/mL for 48h increased apoptosis rate from (4.75 ±1.04)% to (45.61 ±5.22)% with SubG1 (t =7.68,P <0.01 ),and (7.28 ±1.99)% to (57.02 ±3.88)% with Annexin -V /PI(t =11.42,P <0.01).Lovo cells treated with KangAi injection 180μL/mL for 48h increased the expression of Caspase -3 at mRNA level(t =4.06,P <0.05),while Bcl-2 decreased(t =5.69,P <0.01 ).Lovo cells treated with KangAi injection 180μL/mL for 48h decreased the expression of ki -67 from (95.79 ±1.66)% to (65.84 ±4.80)%(t =5.90,P <0.01 ).Conclusion KangAi injection can promote apoptosis and inhibit proliferation of rectal cancer Lovo cells in vitro.

Objective To observe the efficacy and side effects of exemestane in postmenopausal breast cancer patients with bone metastasis.Methods One hundred and ten postmenopausal breast cancer patients with bone metastasis were treated with exemestane 25 mg.Results In the evaluable data from 110 patients,the complete remission(CR)was encountered in 7 cases,partial remission(PR)in 28 cases,with a total response rate of 31.8％ ;Thirty nine patients had stabled diseases for more than 24 weeks.It produced a clinical benefit (CR + PR + SD)over 24 weeks in 74 cases(67.3％).Diseases progressed in 12 of the cases(10.9％).The patients with positive ER and PR status had a higher chance to be benefited from the treatment than those with negative receptor status.The clinical efficacy was not correlated with treatment history,pathological subtypes and bone,liver,lung and lymph node metastasis(x2 =0.045,0.078,0.200,P ＞ 0.05).No severe adverse effects were observed.Conclusion Exemestane is effective to treat bone metastasis of breast cancer with minor adverse reactions and good tolerability.

Objective To investigate the protective effect of retigabine (a M-type potassium channel opener) on brains and its mechanism in male mice after acute cerebral ischemia reperfusion(I/R)injury.Methods Seventy male C57BL/6J mice were randomly divided sham-operated group (n=10),middle cerebral artery occlusion (MCAO) group (n=10) and prevention group (n=50) according to the random number table method;mice in the prevention group were then divided into XE991 (a M-type potassium channel blocker) group,RTG-treatment 0 h group,RTG-treatment 1 h group,RTG-treatment 3 h group,and RTG-treatment 6 h group (n=10).The MCAO models were established by suture method,and reperfusion was performed 90 min after cerebral ischemia.In RTG-treatment groups,a single dose of 10.5 mg/kg RTG was injected at the designated varying time points (0,1,3 and 6 h after the reperfusion);in XE991 group,a single dose of 3.0 mg/kg XE991 was injected after the reperfusion;mice in the sham-operated group and MCAO group received the same volume of saline.Twenty-four h after model making,infarct size was measured by TTC staining.HE staining was used to observe the morphological changes of neurons in hippocampal CA1 regions.The apoptotic neurons level and membrane protein CD40L expression in the ischemic penumbra were detected by TUNEL staining and Western blotting.Results In the sham-operated group,brain tissues had no obvious change,no infarction was observed,there was no CD40L expression,and TUNEL staining positive neurons were hardly found.(1) Cerebral artery territory infarction was visible in the MCAO group and intervention group;however,the infarction volume of the RTG-treatment groups was significantly lower than that in the MCAO group (P＜0.05);the infarction volume of the RTG-treatment 6 h group was increased as compared with that of the RTG-treatment 0 h group,RTG-treatment 1 h group,and RTG-treatment 3 h group,without significant difference (P＞0.05).(2) HE staining showed that hippocampal neurons were obviously swollen and necrotic in the MCAO group and XE991 group,while the pathological damages such as brain edema and neuron necrosis were ameliorated significantly in the RTG-treatment groups.(3) As compared with those in the MCAO group,the number of TUNEL staining positive neurons in the RTG-treatment 0 h group,RTG-treatrnent 1 h group,and RTG-treatment 3 h group and CD40L number in the RTG-treatment 0 h group and RTG-treatment 3 h group were decreased significantly (P＜0.05);as compared with that in the MCAO group,the number of TUNEL staining positive neurons increased significantly in the XE991 group (P＜0.05).Conclusion RTG has protective effect on cerebral I/R,and its mechanism might relate to reducing cell excitability and inflammation,thereby inhibiting cell apoptosis;these protection would be less effective when RTG is used outside a defined critical period of time.

OBJECTIVETo investigate the role of different immunoglobulin- like receptor (KIR)haplotypes in haplo- identical hematopoietic stem cell transplantation (HSCT).

METHODKiller cell KIR genotyping was performed on 468 individuals from 156 unrelated families by PCR-SSP. A total of 624 KIR haplotypes from the parents were used for haplotype analysis. Ninety-two patients received haplo-identical HSCT from one of the parents.

RESULTSThe family study showed segregation of one A haplotype and at least 20 unique B haplotypes. The frequency of haplotype A was 72.92% (455/624). The most commonly observed haplotypes in group B were B1, B2, and B3, present at a frequency of 10.26%, 5.77%, and 4.48%, respectively. Compared to KIR gene matched donors (n=17), grafts from KIR gene mismatched donors (n= 14) had a positive effect on survival after haplo- identical HSCT for AML/MDS patients (OS: 88.2%vs 42.9%,P=0.015; RFS: 88.2%vs 35.7%,P=0.007). No effect was observed for ALL/NHL patients (OS: 76.0%vs 75.0%,P=0.727; RFS: 68.0%vs 65.0%,P=0.866). A significantly lower survival rate was observed for transplants from AA (n=52) and AB1/AB2 donors (n=15), compared to other group Bx donors (n=25) (OS: 53.3%vs 96.0%,P=0.017; RFS: 53.3%vs 92.0%,P=0.019). Meanwhile, the risk of relapse was much higher in AA group (n=52) compared to Bx group (n=40) (25.0%vs 5.0%,P=0.009). A higher risk of TRM was observed in AB1/AB2 group (P=0.012). In addition, transplant from donors carried Cen-B was associated with an increased survival compared with Cen-A homozygous donors (OS: 94.7%vs 68.5%,P=0.036; RFS: 89.5%vs 64.4%,P=0.045).

CONCLUSIONOverall, KIR genotyping and haplotype analyses should be useful for selection of the most optimal donors with favorable KIR gene grafts. KIR gene mismatch donors should be preferred for AML/MDS patients. Selecting donors carried Cen- B and avoiding the selection of donors of KIR genotype AA/AB1/AB2 was strongly advisable for haplo-identical HSCT.

Chronic Disease ; Genotype ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Killer Cells, Natural ; Leukemia, Myeloid, Acute ; therapy ; Neoplasm Recurrence, Local ; Receptors, KIR ; genetics ; Survival Rate ; Tissue Donors

Objective:Establish the decision threshold value of mean fluorescence intensity of anti-human leukocyte antigen(HLA)antibody through statistical analyzing the results of international proficiency testing(PT)organized by American Society for Histocompatibility and Immunogenetics(ASHI).Methods:Single antigen reagent and liquid chip(Luminex)technique were used to detect anti-HLA antibody. A retrospective analysis of the HLA antibody PT results of 55 quality control samples from 11 times organized by ASHI from 2012 to 2019 was reviewed.Results:Among 79 kinds of HLA-I antibodies, 21, 43 and 15 types of HLA-A, B and Cw antibodies were detected respectively, while among 44 kinds of HLA-Ⅱ antibodies, 18, 7 and 19 types of HLA-DRB1, DQB1 and DPB1 antibodies were detected respectively. After analyzing the MFI detection value of different specific antibodies in each PT samples at our laboratory and the coincidence rate of the negative / positive results judged by ASHI through summarizing the results of multicenter participating in the same period, MFI values of HLA antibody were arranged from high to low into the intervals of possible saturation value, positive decision value, positive judgment threshold value, suspicious positive reference value and suspicious negative reference value , according to the coincidence rate of 95%, 90%, 80%, 79%~50% and <50%.Thus, the decision limit value table of HLA specific antibody at our laboratory was established. And 42 kinds of HLA antibody types were detected with complete data.When the MFI values of various HLA-I or HLA-Ⅱ antibodies are found to be 80% or more in the table, it can be used to judge the detection of HLA antibodies. When HLA antibody MFI value reaches the positive decision value, it may have a certain guiding significance for clinical diagnosis and treatment. And when antibody MFI value reaches the saturation value and lies in the suspicious positive or suspicious negative reference threshold, it just suggests that the clinical need for dynamic follow-up of anti-HLA antibody detection.Conclusions:The decision limit value of MFI of laboratory HLA antibody is established based on the international PT experimental results, which is of reference value for the interpretation of experimental results and clinical diagnosis and treatment. A transplant ation center should pay attention to the quality control of comparison test between laboratories in the detection of HLA antibodies.

Objective To investigate the risk factors of acute pancreatitis after endoscopic retrograde chloangiopancreatography (ERCP).Methods Clinical date of 1 041 patients who underwent therapeutic ERCP from January 2016 to August 2017 in department of gastroenterology,Zhongnan Hospital of Wuhan University,were retrospectively studied.Among them post-ERCP pancreatitis developed in 53 patients (PEP group) and the remaining 988 patients were assigned in the non-PEP group.The clinical characteristics,procedures of ERCP,gastrointestinal symptoms and laboratory findings after ERCP were compared between two groups.The SPSS software (version 22.0) was applied to analyze the risk factors of PEP.Results The overall incidence rate of PEP was 5.09％ (53/1 041).There were statistic difference in the number of white cells [8.56(5.43,12.23)× 109/L vs.7.12(5.58,9.20)× 109/L,Z=-2.122] and the ratio of neutrophils [0.82 ± 0.11 vs.0.76 ± 0.12,t=-3.612] after ERCP operation;the incidence of endoscopic sphincterotomy (EST) [60.37％(32/53) vs.43.72％(432/988),x2=5.646] and endoscopic papillary balloon dilation (EPBD)[50.94％(27/53) vs.33.00％(326/988),x2=7.230] during operation,and periampullary diverticula [35.85％(19/53) vs.52.98％(523/988),x2=4.619] (all P＜0.05).Logistic analysis indicated that the ratio of neutrophils was independent risk factor of PEP(OR=1.058,95％CI:1.019-1.098,P=0.003).Conclusion The number of white cells,the ratio of neutrophils after ERCP and endoscopic sphincterotomy,endoscopic papillary balloon dilation and periampullary diverticula are associated with PEP,while the ratio of neutrophils is the independent risk factor of PEP.

Objective: To elucidate the expression levels of key immune biomarkers, phosphate and tension homology deleted on chromosome ten (PTEN) and programmed cell death protein1(PD-1),of different immune tolerance pathway in classic Hodgkin’s lymphoma (CHL) to further determine their clinical role and prognostic significance. Methods: The clinical features and prognostic factors of 56 CHL patients, who were admitted to the TianJin Medical University Cancer Institute from February 2003 to August 2013, were retrospectively analyzed. PTEN and PD-1 protein expression levels were analyzed by immunohistochemistry, Epstein-Barr virus encoded RNA (EBER) was performed by in situ hybridization assay. Correlations between the expression of biomarkers and clinicopathologic parameters were examined and survival analyses were performed. Results: This cohort of 56 CHL patients included 34 males and 22 females with a median age of 25 years (ranged from 7 to 71 years). In a univariate analysis, age≥45, IPS score >2, EBER positive, high expression of PTEN protein conferred inferior 5-year OS and 5-year PFS; In a multivariate model, age≥45, IPS score >2, EBER positive, high expression of PTEN protein were identified as the independent adverse prognostic factors for CHL. Conclusions This study suggested for the first time that PTEN was independent prognostic immune biomarkers in CHL, which provided the novel therapeutic strategy of immune therapy for CHL.

ObjectiveTo observe the effect of Guben Jiedu prescription （GBJ） on the epithelial-mesenchymal transition （EMT） of lung cancer A549 cells and to explore the mechanism based on phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodThe GBJ-medicated serum was prepared. Cell viability was detected by methyl thiazolyl tetrazolium （MTT） assay to screen the optimal doses of GBJ-medicated serum for further experiment. A549 cells were classified into normal serum group， low-， medium-， and high-dose GBJ-medicated serum groups （2.5%， 5%， and 10% GBJ-medicated serum）， PI3K/Akt pathway activator SC79 group， and high-dose GBJ-medicated serum + SC79 group. Cell migration ability was measured by wound-healing assay. The protein expression of E-cadherin， N-cadherin， vimentin， Akt， phosphorylated Akt （p-Akt）， glycogen synthase kinase-3β （GSK-3β）， and phosphorylated GSK-3β （p-GSK-3β） was detected by Western blotting， and the mRNA expression of N-cadherin and vimentin by Real-time PCR. ResultCompared with the normal serum， GBJ-medicated serum （2.5%， 5%， 10%， 20%， 40%） decreased the viability of A549 cells （P<0.05）， and 10%， 5%， 2.5% GBJ-medicated serum was respectively selected for the follow-up experiment. The migration ability of cells in the high-， medium-， and low-dose GBJ-medicated serum groups was lower than that in the normal serum group. The expression of N-cadherin mRNA and Vimentin mRNA in A549 cells in the three GBJ-medicated serum groups was significantly lower than that in the normal serum group （P<0.01）. The protein expression of E-cadherin was higher in the high- and medium-dose GBJ-medicated serum groups than in the normal serum group （P<0.01）. The three GBJ-medicated serum groups showed lower protein expression of N-cadherin， vimentin， p-Akt， and p-GSK-3β （P<0.01） and lower expression of p-Akt/Akt， p-GSK-3β/GSK-3β （P<0.05， P<0.01） than normal serum group. Compared with the SC79 group， the high-dose GBJ-medicated serum group demonstrated high protein expression of E-cadherin （P<0.01） and low expression of N-cadherin， vimentin， p-Akt， p-GSK-3β， and p-Akt/Akt， p-GSK-3β/GSK-3β （P<0.01）. Compared with the high-dose GBJ-medicated serum group， high-dose GBJ-medicated serum + SC79 group showed low protein expression of E-cadherin （P<0.01） and high protein expression of N-cadherin， vimentin， p-Akt， p-GSK-3β， p-Akt/Akt， and p-GSK-3β/GSK-3β （P<0.01）. ConclusionGBJ can inhibit the migration and EMT of lung cancer A549 cells by regulating the PI3K/Akt signaling pathway.

Objective: To analyze family-based haplotype frequencies of HLA-A, -B, -C, -DRB1 and -DQB1 genes and their clinical significance. Methods: The data of HLA genotyping in 3568 families undergoing related haploidentical transplantation between 2012 and 2017 at the First Affiliated Hospital of Soochow University were retrospectively evaluated. The HLA genotyping was performed by PCR amplification with sequence-based typing (PCR-SBT) and sequence-specific oligonucleotide probe (PCR-SSOP) methods. The family genetic analysis and haplotype frequencies were also investigated. Results: All the families were divided into 3 groups, including group1 of 1 422 entire families; group2 of 1 310 patients and either of their parents or one of their children; group3 of 836 patients and their HLA≥5/10 matched sibling donors. In the haplotypes with frequencies greater than 0.1% in group1+ group2, the frequency of A*11∶01-B*40∶01-C*03∶04-DRB1*11∶01-DQB1*03∶01, A*02∶07-B*51∶01-C*14∶02-DRB1*09:01-DQB1*03∶03 were significantly different between group1 and group2 (P=0.029, 0.033) . The frequency of A*11∶01-B*46∶01-C*01∶02∶01G-DRB1*09∶01-DQB1*03∶03 was significantly different between group1 and group3 (P=0.035) . The frequency of A*02∶01-B*40∶01-C*07∶02-DRB1*09∶01-DQB1*03∶03 was significantly different between group1 and group2 (P=0.034) , or group1 and group3 (P=0.034) . The frequency of A*24∶02-B*13∶01-C*03∶04-DRB1*12∶02-DQB1*03:01 was significantly different between group2 and group3 (P=0.046) . Conclusion In this study, we summarize the prevalence of haplotype frequencies in terms of HLA-A, -B, -C, -DRB1 and-DQB1. Based on the database of family haplotype analysis, patients and donor candidates are sorted with matched HLA genotype while unmatched HLA haplotype. Even in patients without entire family information, HLA haplotype analysis assists in choosing the optimal related or unrelated donors.

Immune Checkpoint Inhibitors ; Transcriptome ; Immunotherapy ; Biomarkers, Tumor