Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

Objective:To develop a national secondary reference material of Urea and Creatinine in frozen human serum as a standard for metrological traceability.Methods:According to JJF1343-2012 "General and Statistical Principles for Characterization of Reference Materials" and JJF 1006-1994 " Technical Norm of Primary Reference Material ", the homogeneity, stability, and commutability were evaluated;Using the JCTLM recommended methods, the value of the reference materials was assigned through collaboration with 6 accredited reference laboratories from Guangdong Provincial Hospital of Chinese Medicine, Beijing Aerospace General Hospital, Shenzhen Mindray Bio-Medical Electronics, Maccura Biotechnology, Beijing Leadman Biochemistry, and Zhejiang MedicalSystem Biotechnology. Uncertainty components including inhomogeneity, stability and value assignment were evaluated.Results:The results of one-way analysis of variance of homogeneity for the reference materials showed P>0.05, and the stability evaluation was less than the critical value of the t-test. The measured values were in the 95% confidence interval in the four conventional detection systems for commutability, and the certified values and expanded uncertainties were urea:(14.7±0.3) mmol/L ( k=2),Cr:(313.9±14.5) μmol/L ( k=2). Conclusion:The prepared secondary reference materials of urea and creatinine had promising homogeneity, stability, and commutable, the values of urea and creatinine concentration in reference materials were accurate and reliable.

Objective To express adenosine deaminase protein by molecular cloning technique.MethodsTotal RNA was extracted from human leukocytes and the cDNA was obtained by reverse transcription.Whereupon the cDNA was used as a template to amplify adenosine deaminase gene by polymerase chain reaction (PCR) and then integrated it into three prokaryotic plasmids pET-28 b, pET-32 a (+) and pHSIE.The plasmid with the correct sequencing was transformed into E.coli BL21 (DE3) by CaCl2 method for the protein expression.The expression activity of these fusion proteins were detected by Western-blot and SDS-PAGE, with the optimized expression conditions.Results Complete fusion of target gene and three prokaryotic plasmids was observed through sequencing.The expressed and accurate ADA protein was identified by Westernblot and SDS-PAGE.The optimal expression conditions were observed:the protein expression would be induced with 0.4 mmol/L IPTG and incubated at 16℃for 24 hours.Conclusion The prokaryotic vectors of adenosine deaminase (BL21+pET-28 b+ADA, BL21+pET-32 a+ADA, BL21+pHSIE+ADA) were successfully constructed and efficiently expressed.

Objective To explore the significance of combined detection of D-Dimer,cardic troponin I(cTnI)and N-terminal pro-brain natriuretic peptide(NT-ProBNP)in acute coronary syndrome and the correlation between them.Methods 143 patients with acute coronary syndrome were selected as the observation group,and 40 CAG negative people as the control group.The difference between the two groups was compared and the correlation was analyzed.According to different diagnostics,patients in the observa-tion group were separated into 3 groups,unstable angina pectoris,non ST segment elevation myocardial infarction and ST segment elevation myocardial infarction.The correlation of D-Dimer,cTnI and NT-ProBNP with the severity of coronary artery disease was analyzed.Results The levels of D-Dimer,cTnI and NT-ProBNP in the observation group were higher than those in the control group,and the difference was statistically significant(P < 0.05 ).The severity of coronary artery disease had positive correlation with the test results (P <0.05).Conclusion The combined detection of D-Dimer,cTnI and NT-ProBNP could help to diagnose the acute coronary syndrome and the test result has a positive correlation with the severity of acute coronary syndrome.

OBJECTIVETo establish of blood beta hydroxybutyrate (βOHB) threshold for diagnosing type 2 diabetes ketoacidosis (DKA) and explore the relationship between βOHB levels and the severity of DKA.

METHODSCorrelation analysis was performed between serum βOHB and [HCO(3)] in type 2 diabetic patients admitted in the emergency department in the past year. Regression equation was used to calculate the concentration of βOHB corresponding to a [HCO(3)] level of 18.0, 15, and 10.0 mmol/L, and βOHB concentration corresponding to a [HCO(3)] level of 18.0 mmol/l was used as the DKA diagnostic threshold.

RESULTSThe serum βOHB level and [HCO3] concentration showed a good correlation (R²=0.7023, P<0.001). βOHB concentrations that corresponded to a [HCO(3)] level of 18.0, 15, and 10.0 mmol/L were 3.0, 4.70, and 7.5 mmol/L, respectively, in accordance with the severity of DKA. Combined with the blood glucose concentration ≥ 13.9 mmol/L, a blood βOHB≥3.0 mmol/L showed a sensitivity of 99%, specificity of 86%, and total effectiveness of 92.81% for diagnosing DKA.

CONCLUSIONA serum βOHB level above 3.0 mmol/L can be used as the diagnostic threshold of DKA. βOHB can serve as an index for assessing the severity of DKA.

3-Hydroxybutyric Acid ; blood ; Diabetes Mellitus, Type 2 ; blood ; Diabetic Ketoacidosis ; blood ; diagnosis ; Humans ; Sensitivity and Specificity ; Severity of Illness Index

Objective To Evaluate the performance of serum adenosine deaminase assays of different manufacturers and explore the approach for harmonization of test results.Methods It was evaluated the indice including the limit of blank ,precision,linearity range and reference interval of 10 test systems.It was as the reference system by Mindray test system to evaluate the comparability and the difference of ADA results among 10 different systems.The evaluation was performed before and after calibration by a selected fresh serum assigned by the reference system.A commercial calibrator of the minimum matrix effect was selected from 8 different calibrators as the long-term calibrator to harmonize the ADA results of 10 systems.Results The results of LoB were 0.1-6.3 U/L,respectively.The within-run CVs and total CVs of 10 systems were all less than 5%and actual linearity ranges were conformed to claims of manufacturers.After calibration with fresh serum calibrator ,the averaged difference of 10 test systems was reduced from 14% to 3.0%, and the average difference was 1.8% after calibration with long-term calibrator.The common reference interval of all test systems was 5-24 U/L identically.Conclusions The comparability of ADA measurements can be improved by using a common human serum calibrator and the commutable commercial calibrator.And it is necessary and feasible to develop the standardzation of ADA.

Objective To investigate the effects of interleukine-22 ( IL-22 ) on the expression of interleukin-6 (IL-6) by rheumatoid arthritis synovial fibroblasts (RASF), and to analyze their association with IL-17+CD4+T (Th17) cells differentiation.Methods RASF were isolated from six patients with rheu-matoid arthritis ( RA) and cultured in vitro.The expression of IL-6 at mRNA and protein levels by RASF were detected by qRT-PCR analysis and ELISA after treatment with different concentrations of IL -22 for dif-ferent periods of time.Anti-IL-22R1 blocking antibody and inhibitor assay were used to analyze the specific receptor and its downstream signaling pathways associated with IL-6 production.IL-22 pre-treated RASF and CD4+T cells were co-cultured for 3 days in the presence or absence of anti-IL-22R1 or anti-IL-6 to measure the percentage of Th 17 cells by flow cytometry .Results The expression of IL-6 by RASF was increased up-on IL-22 stimulation in a dose and time dependent manner (P<0.05), and that was closely related to IL-22R1 and its downstream signaling pathways of p38 and JAK2 (P<0.05).Co-culturing CD4+T cells with RASF and Transwell system indicated that the percentage of Th 17 cells was increased in IL-22 pre-treated group as compared with that in IL-22 untreated group , but it could be down-regulated by either blocking IL-22R1 or IL-6.Conclusion IL-22 promoted the expression of IL-6 by RASF and further enhanced Th 17 dif-ferentiation.Neutralizing IL-22 in synovium of patients with RA might be an effective therapeutic strategy for RA treatment.

Objective To explore the effects of Tongfu Huayu Daotan Decoction (通化阏导痰汤)on the serum concentration of S-100β protein, neuron-specific enolase(NSE) and Prognosis in patients with severe craniocerebral injury.Methods Sixty patients with severe craniocerebral injury were randomly divided into a a'eatment group and a control group.The treated group was treated with Tongfu Huayu Daotan Decoction plus conventional treatments including dehydration,antibiotics, organ functional support, nerve nutrition, prevention of complication, etc.; the control group was treated with conventional treatments alone. The concentration of serum S-100β protein and neuron-specific enolase(NSE) in plasma at admission and at24, 36, 72huors, and 5, 7 days after treatment were determined respectively; the Glasgow outcome scale (GOS)and neurological deficits scoring at 2weeks and 4weeks after hospitalization were compared to observe the efficacy of the patients. Results The concentration of serum S-100β protein and neuron-specific enolase (NSE) in plasma at previous treatment and at 24, 36 hours after the treatment had no statistical difference in the two groups(P＞0.05 ), The concentration of serum S-100β protein and neuron-specific enolase (NSE) in plasma at 72huors, 5 and 7 days after the treatment in the Tongfu Huayu Daotan Decoction group were lower than those in the control group, the differences being significant (P＜0.01). The Glasgow outcome scale (GOS) and neurological deficits scoring at 2weeks and 4weeks after the treatment in the Tongfu Huayu Daotan Decoction group were significantly nigher than those in the control group, the differences being significant (P＜0.01) .Conclusion Tongfu Huayu Daotan Decoction can alleviate the plasma concentrations of S-100β protein and neuron-specific enolase (NSE) in patients with severe craniocerebral injury and markedly improve the clinical therapeutic effects. Combined Tongfu Huayu Daotan Decoction and western medicine can significantly reduce mortality and improve the Glasgow outcome scale (GOS), neurological deficits scoring and therapeutic effect.

AIM: To investigate the potential of murine epidermal stem cell (ESC) differentiation after seeded in a biodegradable carrier and implanted subcutaneously into syngeneic recipient mice. METHODS: ES cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a polyglycolic acid (PGA) net containing collagen gel, functioning as a cell carrier, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. RESULTS: The ESCs kept alive in the implant when observed under a fluorescent microscopy 3 weeks or longer after implantation, and could differentiate into hair follicle - like structure,glandular structure, and gave rise to additional structures displaying features resembling native dermis. No apparent rejection or severe side effects were observed at least 10 weeks post- implantation. CONCLUSION: It is feasible to use these ESCs as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.

Objective To evaluate the efficacy of standard large trauma craniectomy to treat the patients with severe bump brain injury. Methods Thirty-two bump brain injured patients with cerebral hernia were treated with inside and outside decompression of standard large trauma craniectomy compared with 24 cases of routine temporoparietal craniectomy between Oct 2001 and Aug 2004.Results In standard large trauma craniectomy group,21 cases got good recovery(65.6%) and 5 were dead(15.6%).In routine temporoparietal craniectomy group,9 cases got good recovery(37.5%) and 10 were dead(41.7%).There was significant difference between the two groups in good recovery and mortality(P