Objective To summarize and analyze the efficacy and influencing factors of second allogeneic hematopoietic stem cell transplantation (allo-HSCT) for acute leukemia relapsing after the first allo-HSCT. Methods Clinical data of 17 patients with acute leukemia who underwent second allo-HSCT at Peking University First Hospital from January 2005 to December 2024 were retrospectively analyzed. Results Among the 17 patients, 7 achieved long-term disease-free survival after second transplantation. The median progression-free survival after successful second transplantation was 7 months (range 8 days to 69 months). The relapse fatality was 24%, and the transplant-related fatality was 35%. Conclusions Second transplantation is an effective treatment for relapsed and refractory acute leukemia, but the relapse fatality and transplant-related fatality remain high. Patient age, time of relapse after the first transplantation and disease status before second transplantation are all factors that affect the efficacy of second transplantation. Younger age, late relapse and complete remission of disease before second transplantation are all beneficial for long-term disease-free survival after second transplantation.

Objective:To explore the safety of laparoscopic antegrade modular pancreatosplenectomy (L-RAMPS) through vascular axis approach in the treatment of pancreatic body and tail adenocarcinoma.Methods:The clinical data of 12 patients with pancreatic body and tail adenocarcinoma undergoing L-RAMPS in Department of Hepatobiliary Pancreatic Surgery, Anhui No.2 Provincial People's Hospital from April 2021 to December 2022 were retrospectively analyzed, including eight males and four females, aged (65.8±11.6) years. Data regarding operative time, intraoperative blood loss, anal exhaust time, postoperative hospital stay, lymph node dissection, pathology, and postoperative complications, and survival were analyzed.Results:The procedures were successfully completed in all 12 patients. Eight patients underwent anterior L-RAMPS, four underwent posterior L-RAMPS. In one patient laparoscopic procedure was almost completed, but eventually conversed to open surgery due to vascular invasion. The operative time was (221.5±21.7) min, the intraoperative blood loss was (224.1±125.3) ml, the anal exhaust time was (3.5±1.0) d, and the postoperative hospital stay was (10.0±3.9) d. All patients underwent R 0 resection, and (15.1±3.7) lymph nodes were dissected. Positive lymph nodes were confirmed in four patients. Six patients had postoperative pancreatic fistula. The patients had been followed up for a median time of 9.5 (3.2-15.0) months, and three patients died. Conclusion:The vascular axis approach could optimize the L-RAMPS surgical approach and improve surgical safety.

Lung cancer is one of the most prevalent malignancies with the highest morbidity and mortality rates worldwide. In recent years, with the development of immune-oncology research and several therapeutic antibodies have reach the clinic, many breakthroughs have been made in immunotherapy. The advent of immunotherapy has revolutionized the treatment of NSCLC, but the response and durable clinical benefit are only observed in a small subset of patients. Therefore, strategies to screen the potential beneficial population and improve the efficacy of immunotherapy remain an essential topic. In the current article, the author review the biomarkers that have potential to better predict responders to immunotherapy and to provide ideas for the clinical application of immunotherapy.
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B7-H1 Antigen ; Biomarkers, Tumor ; Carcinoma, Non-Small-Cell Lung/therapy* ; Humans ; Immunotherapy ; Lung Neoplasms/therapy*

Objective:To discuss the problems existing in the implementation of the Chinese health industry standard WS 523-2019 by testing SPECT device.Methods:Under the WS 523-2019 standards, a total of 10 SPECT devices were tested with regard to their SPECT reconstructed spatial resolution (SRSR), system planar sensitivity (SPS), system spatial resolution (SSR), whole body system spatial resolution (WSSR), intrinsic uniformity (IU), intrinsic count rate performance (ICR), intrinsic spatial resolution (ISR) and intrinsic spatial linearity(ISL).Result:Under the requirements of WS 523-2019 standards for qualified limits, there are 3 devices with ISL unqualified and the rest of the performances qualified. The new standards basically can meet the clinical requirements and reflected the overall performance of SPECT.Conclusions:The distance between the radiation source and the surface of the detector has great influence on the spatial resolution.In the measurement of ISL, there must be a lead grid separately in the x and y directions. The lead grids with the parallel slits shall be positioned on the detector with the center slit centered on the detector. It is suggested to add rotation center in the new standards.

Objective:To construct an expression system of lentivirus vector encoding epidermal growth factor receptor-specific chimeric antigen receptor (EGFR-CAR) and programmed cell death ligand-1 (PD-L1) antibody.Methods:Human PD-L1-Fc protein was used to immunize BALB/c mice. Cell-fusion and subcloning were performed to screen stable hybridoma strains with high secretion of PD-L1-specific antibodies, which were identified by both ELISA and Western blot. The activity of the antibodies in blocking the binding of programmed cell death-1 (PD-1) to PD-L1 was determined by fluorescence-activated cell sorting (FACS). Antibody affinity was analyzed by Fortebio Octet96. A single-chain variable fragment (scFv) was further constructed after antibody full-length sequencing and humanization using CDR grafting method. Meanwhile, the genes encoding the light and heavy chain variable regions (VL and VH) were cloned from a hybridoma secreting antibody against human EGFR by 5′ RACE technology to construct scFv gene. The expression of scFv was confirmed using pcDNA3.1 vector. EGFR-CAR containing CD137 intracellular function domain and PD-L1-scFv was ligated using 2A gene. The synthetic single molecule was cloned into pLVX-EF1a-IRES-ZsGreen1 lentivirus expression vector, and then transfected into 293T cells using Lenti-X Packaging Single Shots (VSV-G) to prepare infectious virus. Expression of CAR on cell surface and the soluble form of PD-L1-scFv in the supernatant of transfected 293V cells were detected by FACS and ELISA.Results:A PD-L1 antibody named 11E3 with high ligand-receptor blocking performance was obtained. The humanized antibody showed a stable affinity (2.67×10 -10 mol/L) after directly grafting the mouse CDRs (CDR1, CDR2 and CDR3) to human frameworks. EGFR-scFv was effectively expressed in a form of Fc-fusion. Secretory CAR (CTZ0431-1) and membrane CAR (CTZ0431-2) expression plasmids were constructed using lentivirus vector containing EGFR-CAR and PD-L1-scFv. The infection efficiency in 293V cells was around 10%. EGFR-scFv on the cell membranes and PD-L1-scfv in the culture supernatants were detected after 293V cells were infected with CTZ0431-1. EGFR-scFv and PD-L1-scfv were expressed on the cell membranes of 293V cells infected with CTZ0431-2. The expression rate of CAR in LV-CART46407-1-transfected activated T cells was 39.3%. Conclusions:The lentivirus vectors co-expressing EGFR-CAR with moderate binding affinity and PD-L1-scFv with high binding affinity were successful constructed, which provided an essential tool for investing EGFR- and PD-L1 double targeted CAR-T cell therapy against solid tumor.

Objective To analyze the epidemiology and clinical characteristics of patients with Brucella endocarditis in order to improve the understanding and treatment of Brucella endocarditis.Methods The clinical data including demographic characteristics,clinical features,laboratory data,echocardiography,treatment and clinical outcome of 6 patients with Brucella endocarditis were collected and analyzed.Results In our database consisting of 211 brucellosis patients with positive blood culture,6 patients (2.8％) with Brucella endocarditis were selected from May 2007 to December 2016 in Peking Union Medical College Hospital.The age range was 25-55 years,among whom 5 patients were men.The majority were farmers and all patients reported the history of closely contact with livestock.All of them had fever and 3 of them had arthralgia.Four patients had aortic involvement (4/6) and 2 of them presented with symptoms of left ventricular failure at admission.Blood culture of Brucella and serum agglutination test of Brucella were positive in all of them.Combined antibiotics treatments were given to all patients.Valve replacement surgery was performed in four patients,among them,three patients well recovered,one lost.Among all the patients,four were followed up continually and other two were lost.Conclusions Brucella endocarditis has a predominance of aortic involvement and is prone to left ventricular failure.Early antibiotic treatment combined with valve-replacement surgery is effective to improve the prognosis.

In this study, hollow fiber membrane extraction combined with ambient ionization mass spectrometry ( AMS) was developed for the simultaneous determination of 7 perfluorinated compounds ( PFCs) in aqueous solution, including perfluoroheptanoic acid ( PFHpA ) , perfluorooctanoic acid ( PFOA ) , perfluorooctane sulfonate acid ( PFOS ) , perfluorononanoic acid ( PFNA ) , perfluorodecanoic acid ( PFDA ) , perfluoroundecanoic acid ( PFUdA) , and perfluorododecanoic acid ( PFDoA) . PFCs were detected in negative ion mode using selective reaction monitoring ( SRM) mode. The extraction time and the pH value of extraction solution were optimized. 13 C4-PFOS and 13 C4-PFOA were used as internal standards for quantitative analysis. The method showed good linearity with correlation coefficient values ( r2 ) greater than 0. 991 for the seven target PFCs. With the exception of PFHpA, the limit of detection ( LOD) for other six PFCs was within ranges from 0. 8 to 2. 7 ng/L while the limit of quantitative (LOQ) was from 2. 7 ng/L to 8. 9 ng/L. The enrichment factor of five PFCs was more than two hundred. The developed method was applied to detect the seven PFCs in tap water and Pearl River water, and they were all not detected. The recoveries were within the ranges of 88. 5%-108. 3% and 94. 2%-116. 7% when 40 ng/L and 400 ng/L PFCs were spiked into tap water, respectively. In terms of the Pearl River water, the recoveries were within the ranges of 75. 0%-102. 6% and 81. 2%-97. 6% when 40 ng/L and 400 ng/L PFCs were spiked, respectively.

Objective To investigate the effect of intrathecal hyperbaric bupivacaine on spinal cord neurons apoptosis in diabetic neuropathic rats.Methods Thirty-two healthy male Sprague-Daw-ley rats weighing 220-300 g,8 normal rats randomly served as control group (group C),the other rats were intraperitoneal injection 1% streptozotocin (STZ)60 mg/kg to induce diabetic neuropathic (DN),and last induced thirty-seven diabetic neuropathic rats.group C and diabetic neuropathic rats administer intrathecal catheter,respectively.Twenty-four rats in which DN was successfully intrathe-cal catheter were randomly divided into 3 groups (n=8):hyperbaric bupivacaine group (group HB), isobaric bupivacaine group (group IB),glucose group (group G).Hyperbaric bupivacaine 10 μl were injected intrathecally in groups C and HB respectively,isobaric lidocaine 10 μl were injected intrathe-cally in group IB,10% glucose 10 μl were injected intrathecally in group G,once daily for 3d.After rats each administration 2 min,motor block duration were recorded;The paw withdrawal threshold to von Frey filament stimulation (PWT)were measured before induced diabetes model (T1 ),before in-jected intrathecally (T2 ),after 30 min administered 1 d (T3 ),2 d (T4 ),3 d (T5 )and end administered 4 d (T6 ).All rats were sacrificed at T6 and their lumbar intumescential spinal cord tissue were re-moved for microscopic examination.And using TUNEL assay to measure spinal neuronal apoptosis. Results PWT was lower at T2-5 in groups HB,IB and G comparing with T1 (P <0.05 ).Comparing with group C,the motor block duration was significantly prolonged(P <0.05)and spinal cord neuro-nal apoptosis cells were increased(P <0.05)in group HB.Comparing with group IB,the motor block duration was significantly prolonged(P <0.05)and spinal cord neuronal apoptosis cells were increased (P <0.05)in group HB,too.PWT was increased at T6 in group HB at T2-T5 (P <0.05).Group G did not appear motor block and spinal cord neuronal apoptosis.Conclusion Intrathecally hyperbaric bupi-vacaine can promote spinal cord neuronal apoptosis in diabetic neuropathic rats.

Objective To determine whether regulatory T cells(Tr)are increased in patients with tuberculosis and whether they are associated with its immunopathology.Meantime,to investigate the possibility of tuberculosis(TB)as a model for studying Tr functions.Methods The lymphocyte subsets were isolated from peripheral blood mononuclear cells by sorting with flow cytometry.Total cellular RNA was extracted and RT-PCR was performed to detect the Foxp3 mRNA in purified CD3+CIM+T cells,CD3+CD8+T cells and non-CD3+CD4+CD8+T cells.Using FACS analysis.we further investigated the distribution of Foxp3+ population in CD4+ CD25+T cells.Finally,we compared the percentage of CD4+CD25highFoxp3+T cells present in 51 active patients with tuberculosis and 40 uninfected healthy control subjects by FACS.The detection of Tr infiltration of Foxp3+ cells were performed with immunohistochemistry(IHC)method on tuberculosis pathological sections.Results Foxp3 was specific expressed in CD3+CD4+T cells,either in tuberculosis patients or healthy control subjects.Foxp3+ T cells took about 85%fraction of CD4+ CD25highpopulation.We used CD4+CD25high Foxp3+as a detective markers for Tr in the FACS analysis.The results showed that patients with active TB had a 4.4 fold higher percentage within the CD4+T cells in peripheral blood compared to healthy control group(modian,1.01%vs 0.23%,P<0.01).Much higher frequency of Tr were found along with T cells infiltration at the tuberculosis pathological tissues.A few individuals that we can followed indicated the expanded Tr was declined after curative treatment with operation.Conclusion Tr cells are increased in tuberculosis patients and closely correlate with its immunopathology.Tuberculosis should be a valuable model for Tr functional study.

Objective To explore the application of interferon(IFN)response-related genes in predicting the outcome of antiviral treatment in chronic hepatitis C. Methods SuperArray microarray was used to detect the expression of IFN response-related genes in peripheral blood monocytes(PBMC)from chronic hepatitis C patients before treatment. SPSS 12.0 was used for statistical analysis. Results Ten patients were classified as rapid responders(RVR)and seven patients as non-RVRs according to the serum HCV RNA level after 4 weeks of treatment in 17 patients. Compared with healthy controls, nine differentially expressed genes were found in RVR patients, one up-regulated and eight down-regulated; eighteen differentially expressed genes were found in N-RVR patients, all down-regulated. Five differentially expressed genes were found between the patients with RVR and N-RVR: four up-regulated genes were PRKCZ, PRKRA, IRF5 and TNFSF10(t =5.44, 3.13, 5.24 and 2.30, P=0. 000, 0.010, 0.005 and 0. 044); one up-regulated gene was IFIT5(t = 2.43, P = 0. 035). Of seventeen patients, 12 were HCV genotype 1b, 5 were HCV genotype 2a. Compared with HCV2a, IFI6 and IFI44 gene in HCV1b were downregulated(t = 2.42 and 2.45, P = 0. 038 and 0. 033). Conclusions The expression of IFN responserelated genes is associated with response to IFN treatment. HCV genotype 1 b is more successful in inducing the down-regulation of IFN response-related genes than that of HCV genotype 2a, thus leading to the resistance to IFN.