Objective To evaluate the analytical performance of ARCHITECT-i2000SR chemiluminescence analyzer in detec-tion of HBsAg,HIV-antibody,TP-antibody and HCV-antibody.Methods Validated the precision,carryover,ference interval of alpha-fetoprotein and linearity performance of ARCHITECT-i2000SR.The same specimens were tested by both CMIA and ELISA,and the results were compared and analyzed.Results The precision,carryover and ference interval of alpha-feto-protein of HBsAg,HIV-antibody,TP-antibody and HCV-antibody were within the range provided by the manufacturer for ARCHITECT-i2000SR.The theoretical and measured values of HBsAg were:Y = 1.102X - 6.678 6 (r = 0.995 5,P <0.05),the range of linearity 1.08~208.6.The sesult was very good for the four blood index by both methods.Conclusion The basic performances of ARCHITECT-i2000SR were consistent with the data provided by the manufacturer,so it canbe used to inspect the clinical samples and the sasults were credible.

BACKGROUND:Stromal cel-derived factor-1 (SDF-1) is one of the most powerful chemokines in myocardial infarction region and plays a particularly pivotal role in the homing of stem cels to an injured myocardium and promoting angiogenesis. On the other hand the microbubble and acoustics active substances carrying targetable ligands can be prepared into targeted ultrasound contrast agents that can be combined with living cels used for molecular imaging. The key of ultrasonic molecular imaging is to find imaging targets, and to successfuly prepare targeted ultrasound contrast agent which can be combined with the imaging target specificaly and efficiently. OBJECTIVE:To prepare and evaluate targeted microbubble contrast agents with SDF-1 monoclonal antibody. METHODS:Targeted microbubble contrast agent with SDF-1 monoclonal antibody was prepared using the biotin-streptavidin method. The physiochemical properties of targeted microbubble contrast agent were evaluated by appearance, pH, particle diameter, optical and fluorescence microscope and flow cytometry test. Four minipigs underwent ligation of the left anterior descending coronary artery to complete the establishment of acute myocardial infarction model, and another two minipigs were subject to thoracotomy but no ligation of the coronary artery. Then, al animals were injected with microbubble contrast agents. The stability of microbubbles was assessed by immunofluorescence testin vivo. RESULTS AND CONCLUSION:SDF-1 and microbubbles were combined by biotin-streptavidin method.In vitro appearance of the contrast agent was translucent yelow or green, and stratified after standing. pH vaule was 7.02±0.12 for non-targeted contrast agent and 6.10±0.19 for targeted microbubble contrast agent. Under the fluorescence microscope, the distribution and size of targeted microbubbles were uniform, and the microbubbles were surrounded by bright and ring shaped green fluorescence that had no changes after highly shaking. The diameter of microbubbles was (2 422.62±238.82) nm after carrying the SDF-1 antibody. Flow cytometry results showed that the carrying rate of targeted contrast agents was stable in different periods.In vivo test showed that targeted microbubbles gathered in vascular endothelial cel surface after acute myocardial infarction. These findings indicate that the targeted microbubble contrast agent carrying SDF-1 monoclonal antibody prepared by biotin-streptavidin method can be combined with vascular endothelial cels, and the binding rate is high and stablein vitro.

OBJECTIVEThe purpose of this study was to evaluate the advantages and disadvantages of vascularized free fibula flap as a new method for mandibular reconstruction.

METHODS25 cases (17 male to 8 female) who have received mandibular reconstruction with free vascularized fibular flaps in our hospital were studied retrospectively. The average length of the fibula grafts is 10.0 cm (range from 5.5 to 16 cm). 3 cases received primary insertion of osteointegrated dental implants into the free fibula flap, and all these 5 implants survived.

RESULTSAll flaps except 1 were viable. 62% of the cases took normal diet postoperatively, and the remainder took soft diet as well. All patients spoke clearly. No ankle unstability was reported. And the aesthetic assessments in all patients were good or fair.

CONCLUSIONVascularized free fibular flap takes its distinct advantages to other autogeneous free bone flaps and is confirmed to be one of the optimal methods for mandible reconstruction by our study.

Adolescent ; Adult ; Aged ; Fibula ; blood supply ; transplantation ; Follow-Up Studies ; Graft Survival ; Humans ; Male ; Mandible ; surgery ; Mandibular Neoplasms ; rehabilitation ; surgery ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps

OBJECTIVETo assess the association of 8 single nucleotide polymorphisms (SNPs) from chromosomes X and 20 with androgenetic alopecia among ethnic Han population from Yunnan province.

METHODSAn eight-SNP co-amplification protocol was developed for the genotyping with a SNaPshot platform. A case-control study was carried out for the 8 SNPs from chromosomes X and 20 in 115 androgenetic alopecia cases and 125 healthy controls. Statistical analysis was conducted with SPSS17.0, Haploview4.2, SHEsis and MDR software.

RESULTSNo association was found between the two groups with regard to the 4 SNPs located on the X chromosome. The genotypic frequencies of rs2180439, rs913063 and rs1160312 were significantly different between the two groups (P < 0.05). The frequency of T allele of rs2180439 was significantly higher in the case group (P < 0.05). The frequencies of A alleles of rs913063 and rs1160312 were significantly higher in the case group (P < 0.05). The haplotypes of C-T-C-G, T-C-C-G and T-T-A-A based on rs6137444-rs2180439-rs913063-rs1160312 showed significant difference between the two groups (P <0.05). rs6137444, rs21804393 and rs1160312 have a strong association with androgenetic alopecia.

CONCLUSIONThe 4 SNPs located on chromosome X were all monomorphic among ethnic Hans from Yunnan. The rs6152, rs16990427, rs1352015, rs1385699 SNPs located on chromosome 20 are associated with androgenetic alopecia in the same population. Individuals with T allele of rs2180439 and A allele of rs913063 and rs1160312 are more likely to develop androgenetic alopecia.

Adult ; Alopecia ; genetics ; Case-Control Studies ; China ; ethnology ; Chromosomes, Human, Pair 20 ; Chromosomes, Human, X ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

To retrospectively analyze the efficacy and safety of modified cell infusion method in reducing the incidence of febrile non?hemolytic transfusion reaction (FNHTR). Methods A total of 69 patients were enrolled in the clinical trial of CD19 chimeric antigen receptor T (CAR?T) cell treatment from February 2017 to October 2018. Study group received the modified cell infusion method, that 1×106 CAR?T cells were re?suspended in 2 mg human serum albumin with total volume of 20 ml and injected intravenously. The control group was intravenously administrated with CAR?T cell in 100 ml normal saline. The incidence of FNHTR, cytokine releasing syndrome (CRS) grade, cytokine level and efficacy were compared. Results (1)The incidence of FNHTR in the study group was 21.1%, significantly lower than that in the control group (71%)(P=0.000). (2)There was no statistical difference in cell proliferation between the study group and the control group on day 4, 7, 14 and 21 after CAR?T cell infusion (P=10.223, 3.254, 5.551, 7.605). (3)There was no statistical difference in CRS grading between the study group and the control group (P=0.767). There was no statistical difference in the levels of interleukin 2 receptor (IL?2R), IL?6, tumor necrosis factor (TNF)?α between the two groups. (4)The C?reaction protein (CRP) level of the study group was lower than that of the control group on day 4 and 7 (P=0.026, 0.007). (5)There was no statistical difference of response rates in acute lymphocytic leukemia (ALL) and non?Hodgkin lymphoma (NHL) patients between the two groups (PALL=0.842; PNHL=0.866). Conclusion The modified cell infusion method in CD19 CAR?T cell treatment reduces the incidence of treatment?related FNHTR. It does not affect the proliferation of CAR?T cells in vivo, the grading of CRS and the response rates.

Objective To investigate the clinical value and experience of transperitoneal and retroperitoneal robot-assisted partial nephrectomy for renal hi1 ar tumors.Methods We evaluated 48 patients who had partial nephrectomy for renal hilar tumor by robotic surgical syestem from January 2013 to March 2017.In those cases,35 were male and 13 were female,with an average age of 57.3 (range from 41 to 75 ),27 cases were ventral tumor and 21 cases were dorsal tumor.3 cases were totally confined to the renal parenchyma,the other 45 cases were partially confined to the renal parenchyma.18 cases were performed surgery by retroperitoneal route,the rest 30 cases were performed by peritoneal route.Results A total of 48 patients underwent successful robotic partial nephrectomy for renal hilar tumors.The mean warm ischemia time was 22 minutes (range from 16 to 33 minutes) and the mean estimated blood loss was 88 md (range from 50 to 350 ml).No bleeding-related complications were found.Histopathology confirmed 39 cases of ccRCC,7 cases of angioleiomyolipoma,2 cases of renal oncocytoma.There was one case in this review was positive surgical margin (2.1％) and found no sign of recurrence during the short term post-operation follow-up.All cases in this review are following up after surgery to date from 2 months to 4 years,no cases of tumor recurrence or metastasis were found.Conclusions The application of transperitoneal and retroperitoneal RAPN is the effective and safe way for renal hilar tumor resection,and it has a clear advantage of renal surgical incision stitching and tumor complete resection.The choice of surgical approaches depends on the size and location of tumor and the clinical experience of the surgeon.

Objective: To investigate the efficacy and safety of CD19 chimeric antigen receptor T (CAR-T) lymphocytes for the treatment of B cell lymphoma. Methods: A total of 22 patients with B-cell lymphoma from February 1, 2017 to July 1, 2018 were reviewed to evaluate the efficacy and adverse reactions of CD19 CAR-T. Results: Of 22 patients with B-cell lymphoma received CD19 CAR-T cells, the median dose of CAR-T cells was 7.2 (2.0-12.0) ×10⁶/kg. Nine of 12 cases of relapse refractory patients were overall response. Complete remission (CR) occurred in 2 of 12 patients, partial remission (PR) in 7 of 12 patients. The overall response in minor residual disease positive (MRD) group was 8 of 10 patients. CD19 CAR-T cells proliferated in vivo and were detectable in the blood of patients. The peak timepoints of CAR-T cells proliferated in the relapsed refractory and MRD positive groups were 12 (5-19) and 4.5 (1-12) days after treatment respectively, and among peripheral blood cells, CAR-T cells accounted for 10.10% (3.55%-24.74%) and 4.02% (2.23%-28.60%) of T lymphocytes respectively. The MRD positive patients achieved sustained remissions during a median follow-up of 8 months (rang 3-18 months) . None of all the patients relapsed during a median follow-up time of 10 months (3-18 months) . However, 7 PR responders of the relapsed refractory patients maintained a good condition for 1.5-6.0 months. One patient bridged to hematopoietic stem cell transplantation, another one sustained remission for 12 months. Cytokine-release syndrome (CRS) occurred in 14 patients with grade 1-2 CRS in MRD positive group and grade 3 CRS in relapsed refractory group. Conclusions CAR-T cell therapy not only played a role in the rescue treatment of relapsed and refractory patients, but also produced a surprising effect in the consolidation and maintenance of B-cell lymphoma. CD19 CAR-T cells might be more effective in the treatment of MRD positive B-cell lymphoma patients than in the refractory or relapsed cases. High response rate was observed with fewer adverse reactions.

Objective: To Evaluation the effect of PD-1 inhibitor Nivolumab on the proliferation and cytotoxicity of anti-CD19 chimeric antigen receptor T cells (CD19-CAR-T) in vitro. Methods: Five patients with high PD-1 expression in peripheral blood and five healthy volunteers were selected. These peripheral blood mononuclear cells were used as the source of T cells to prepare CD19-CAR-T cells. Different doses (72, 36, 18 μg/ml) of Nivolumab was added on day 8 to the culture medium. Patient T cells incubated with 72 μg/ml Nivolumab and CD19-CAR-T cells of healthy volunteers were used as controls. CCK-8, lactate dehydrogenase (LDH) cytotoxicity assay and ELASA were used to detect the proliferation capacity, the specific cytotoxicity and the inflammatory factor secretion. Results: ①T cells from patients with high expression of PD-1 as the source of CD19-CAR-T cells did not affect transfection rate compared with that of healthy volunteers [(32.80±7.22)% vs (35.10±5.84)%, t=-0.554, P=0.593]. ②Incubation of CD19-CAR-T cells with 72 μg/ml Nivolumab did not affect CD19-CAR-T cell proliferation, but its cytotoxicity was significantly higher than that of CD19-CAR-T cells alone or patients’ T cells +72 μg/ml Nivolumab (all P<0.001), there was no significant difference in the killing activity between the 72 μg/ml and 36 μg/ml Nivolumab treated CD19-CAR-T cells on Pfeiffer cells (P=0.281, 0.267, respectively), and they were all higher than those of 18 μg/ml Nivolumab treated CD19-CAR-T cells (all P<0.001). ③Different doses of PD-1 inhibitor Nivolumab combined with CD19-CAR-T cells does not affect the secretion of IFN-γ and IFN-α (all P>0.05). Conclusion Combination of 36 μg/ml PD-1 inhibitor and CD19-CAR-T cells could reduce the drug toxicity and enhance the cytotoxicity.

Objective: To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells. Methods: The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay. Results: ①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) . Conclusion The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.

Objective: To retrospectively analyze the efficacy and safety of modified cell infusion method in reducing the incidence of febrile non-hemolytic transfusion reaction (FNHTR). Methods: A total of 69 patients were enrolled in the clinical trial of CD₁₉ chimeric antigen receptor T (CAR-T) cell treatment from February 2017 to October 2018. Study group received the modified cell infusion method, that 1×10⁶ CAR-T cells were re-suspended in 2 mg human serum albumin with total volume of 20 ml and injected intravenously. The control group was intravenously administrated with CAR-T cell in 100 ml normal saline. The incidence of FNHTR, cytokine releasing syndrome (CRS) grade, cytokine level and efficacy were compared. Results: (1)The incidence of FNHTR in the study group was 21.1%, significantly lower than that in the control group (71%)(P=0.000). (2)There was no statistical difference in cell proliferation between the study group and the control group on day 4, 7, 14 and 21 after CAR-T cell infusion (P=10.223, 3.254, 5.551, 7.605). (3)There was no statistical difference in CRS grading between the study group and the control group (P=0.767). There was no statistical difference in the levels of interleukin 2 receptor (IL-2R), IL-6, tumor necrosis factor (TNF)-α between the two groups. (4)The C-reaction protein (CRP) level of the study group was lower than that of the control group on day 4 and 7 (P=0.026, 0.007). (5)There was no statistical difference of response rates in acute lymphocytic leukemia (ALL) and non-Hodgkin lymphoma (NHL) patients between the two groups (P_ALL=0.842; P_NHL=0.866). Conclusion The modified cell infusion method in CD₁₉ CAR-T cell treatment reduces the incidence of treatment-related FNHTR. It does not affect the proliferation of CAR-T cells in vivo, the grading of CRS and the response rates.