Objective:This study aims to reveal the special immune infiltrating environment and possible immune escape mechanism of giant cell tumor of bone through single-cell sequencing data.Methods:The fresh samples obtained from 4 patients with primary giant cell tumor of bone from January 2018 to December 2021 were collected, and single-cell transcriptome sequencing was performed on the 10X platform to explore the characteristics and immune environment of giant cell tumor of bone by using t-distributed stochastic neighbor embedding ( t-SNE). The main cell types and signal pathways of immune cell regulation and function in giant cell tumor of bone were observed by cell communication analysis. Results:Cell clustering, the definition of basic cell types, the classification of immune cells, and the mutual regulatory relationship between cell types were analyzed for 35 643 single-cell data from 4 giant cell tumor samples of bone. It was found that giant cell tumor of bone was composed of 9 basic cell types, in which the immune cells were mainly CD8 + T cells (51%) and the non-immune cells were mainly fibroblast like spindle stromal cells (74%). The immune infiltration of giant cell tumor of bone is dominated by cytotoxic CD8 + T cells and lacks exhausted CD8 + T cells. CD4 + T cells are characterized by high expression of immune checkpoint genes CTLA4 and TIGIT. In giant cell tumor of bone, immune cells mainly act on multinucleated osteoclast like giant cells through PARs and CCL signaling pathways, but not stromal cells. Conclusion:This study defined the main cell types of giant cell tumor of bone through single cell sequencing data, and further revealed the composition characteristics of its immune infiltration, and found that the target of its immune cells was mainly multinuclear osteoclast like giant cells, which provided effective information for further understanding the occurrence and development of giant cell tumor of bone.

To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipe-line and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to gen-erate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.

OBJECTIVE： To provide reference for improving the quality standard of Pheretima. METHODS： The contents of hypoxanthine and inosine in medicinal material samples were determined by HPLC. HPLC fingerprint of Pheretima was established according to “Similarity evaluation system for TCM chromatogramtic fingerprint” （2012 edition） software， and similarity evaluation was conducted. The determination was performed on Purospher STAR RP-18 endcapped with mobile phase consisted of methanol-water （gradient elution） at the flow rate of 1 mL/min. The detection wavelength was 248 nm， and the column temperature was set at 30 ℃. The sample size was 20 μL. RESULTS： The results of methodological investigation of content determination showed that the linear range of hypoxanthine and inosine were 1.58-31.6 μg/mL （r＝0.999 9）， 5.52-110.4 μg/mL（r＝0.999 8）， respectively. limits of quantify were 0.316， 0.552 μg/mL， respectively； limits of detection were 0.158， 0.110 μg/mL， respectively； RSDs of precision， stability （24 h） and repeatability tests were all less than 2.0％ （n＝6）. Average recovery rates were 103.0％ （RSD＝1.7％， n＝6） and 101.2％ （RSD＝1.2％， n＝6）， respectively. HPLC fingerprint for 15 batches of samples were established， and 8 common peaks were identified. The similarity of HPLC fingerprint of 14 batches of sample with control fingerprint R was higher than 0.900. CONCLUSIONS： The established method for content determination of hypoxanthine and inosine and HPLC fingerprint of Pheretima are simple， accurate and reproducible， and can be used for quality control of Pheretima.

Objective To analyze the effect and possible mechanism of propofol on proliferation and inva-sion of lung cancer cells. Methods Lung cancer cells were cultured and divided into experimental control group, groupL,group M and group H.The optical density of cancer cell was detected by CCK-8.Meanwhile,the difference of the proliferation of cancer cells in each groupwas determined after down-regulation gene.The change of inhibition rate in four groups was analyzed by Transwell test. The expression of MMP-2 and mRNA in different groups were compared by Western blot and Q-PCR. Results the results of CCK-8 test showed the cancer cell viability in H group was lowest and the inhibition ability increasedafter silencing MMP-2 gene(P<0.05); the invasion inhibition rate of the lung cancer cells treated with propofol was higher(P<0.05), and invasion inhibition rate in each group also rise after silencing MMP-2; Western blot and Q-PCR show that the MMP-2 expression level in group Mand group H were lowest(P<0.05).Conclusion Propofol may inhibit the proliferation and invasion of lung cancer cells and its mechanism may be associated with down-regulation MMP-2,which is expected to inhibit the proliferation of lung cancer invasion.

BACKGROUND: Studies have shown that propofol enables a reduction in the number of adult rat mesenchymal stem cells, while the cell differentiation is also significantly inhibited. OBJECTIVE: To explore whether liver X receptors (LXRs) can reverse the inhibitory effects of propofol on bone marrow mesenchymal stem cells. METHODS: Fifteen healthy C57/BL6 mice were randomized into three groups, 5 of which served as blank control group (intraperitoneally treated with normal saline), 5 as propofol treatment group (intraperitoneally treated with 60 mg/kg propofol), and 5 as propofol + LXRs agonist treatment group (intraperitoneally injected with 10 μL/g LXRs at the 1st day, and then injected with 60 mg/kg propofol at the 2nd day). The mice in the three groups were killed at 1-3 hours after treatment to isolate and culture bone marrow mesenchymal stem cells. Cell counting kit-8 and cloneformation assay were used to evaluate the abilities of cell proliferation and self-renewal; induced differentiation experiments in vitro were used to evaluate the differentiation ability of cells into adipocytes, osteoblasts and chondrocytes; real-time quantitative PCR was used to detect the expression of differentiation related molecules andNotch signal. RESULTS AND CONCLUSION: In the propofol-treated mice, cell viability and clone forming ability as well as adipogenic, osteogenic and chondrogenic differentiation of cells decreased significantly compared with the blank control group (P <0.05), while LXR agonists could reverse these effects significantly (P < 0.05). Notch signal expressions showed no difference among three groups prior to induced differentiation. The expression levels differentiation related molecules downregulated significantly after propofol treatment (P < 0.05), but upregulated significantly after treatment with LXR agonists (P < 0.05). Notch signaling inhibitor treatment could significantly inhibit the multi-directional differentiation of bone marrow mesenchymal stem cells in the three groups. All these findings indicate that activated LXRs can reverse the inhibitory effects of propofol on bone marrow mesenchymal stem cells.

Objective: To study the expression of CD123 in bone marrow (BM) blasts of acute myeloid leukemia (AML) patients to explore the relationship between CD123 expression and therapeutic response and prognosis. Methods: This study retrospectively analyzed expression and distribution of CD123 in BM blasts in 137 cases of newly diagnosed AML (excluded M₃) , CD123 detected by flow cytometry≥20% was defined as positive, including 84 CD123⁺ AML and 53 CD123^- AML, efficacy and prognosis were compared between the two groups. Results: ① Among 137 patients, 84 were in group CD123⁺ (61.3%) , and 53 in group CD123^- (38.7%) . All 137 patients were classified into risk groups based on cytogenetic and molecular biology abnormalities. No significant differences were seen between the three risk groups with regard to their CD123 levels (χ²=0.861, P=0.650) . Compared with CD123^- group, the CD123⁺ group had higher WBC[47.7 (1.0-264.0) vs 22.4 (0.7-211.0) , z=-2.592, P=0.010]. ② The rates of first complete remission (CR1) and recurrence of CD123⁺ group were 54.8% (46/84) and 50.8% (32/63) , respectively; and CD123^- group were 73.6% (39/53) and 41.7% (20/48) , respectively. There was significant difference of CR1 between the two groups (χ²=5.121, P=0.027) , whereas no significant difference of the recurrence rate (χ²=0.911, P=0.340) . ③ The median dutations of OS between CD123⁺ group and CD123^- group were 20.0 (95%CI 13.1-26.9) months vs 44.0 (95%CI 23.6-47.3) months, respectively (χ²=5.874, P=0.015) ; The median durations of DFS were 7.8 (95%CI 1.4-14.1) months vs 18.6 (95%CI 0-39.7) months, respectively, no differences were observed between the two groups (χ²=2.939, P=0.086) . ④ CD123 retained an adverse prognosis value on DFS and OS within the intermediate group and patients ≤ 50 years older. Conclusions CD123 widely expressed in AML patients, which was an independent risk factor for CR1 and OS, which implicating its important role in evaluating the induction chemotherapy response and prognosis of AML.

Objective To investigate the role of PI3-kinase-Akt-endothelial nitric oxide synthase (PI3KAkt-eNOS) signaling pathway in the attenuation of myocardial ischemia-reperfusion (I/R) injury by sevoflurane postconditioning in rats.Methods Fifty healthy male Wistar rats weighing 250-280 g aged 2-3 months were randomly divided into 5 groups ( n =10 each):sham operation group (group S),I/R group,sevoflurane postconditioning group (group Spo),sevoflurane postconditioning + dimethyl sulfoxide (DMSO) group (group Spo + D),and sevoflurane postconditioning + LY294002 (a specific PI3K inhibitor) group (group Spo+ L).I/R was produced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in anesthetized rats.In group Spo,sevoflurane was inhaled for 5 min after the end-tidal concentration reached 2.5％-3.0％ at 1 min before reperfusion.In group Spo + L,LY294002 0.3 mg/kg in 0.02％ DMSO was injected intravenously at 5 min before reperfusion,and then sevoflurane postconditioning was performed.In group Spo + D,0.02％ DMSO equal to the volume of LY294002 was injected intravenously at 5 min before reperfusion,and then sevoflurane postconditioning was performed.Arterial blood samples were taken at 120 min of reperfusion for determination of the levels of creatine kinase isoenzyme MB (CK-MB),lactate dehydrogenase (LDH) and cardiac Troponin Ⅰ (cTnI).The myocardial infarct size (IS) and area at risk (AAR) were measured and IS/AAR ratio was calculated.The rats were sacrificed at 120 min of reperfusion and the myocardial tissues in the area at risk were taken for determination of the expression of Akt,phosphorylated Akt (p-Akt),eNOS and phosphorylated eNOS (peNOS) by Western blot.The ratios of p-Akt/Akt and p-eNOS/eNOS were calculated.Results Compared with group S,the levels of CK-MB,LDH and cTnI,IS/AAR ratio,p-Akt/Akt ratio and p-eNOS/eNOS ratio were significantly increased in the other groups ( P ＜ 0.05 or 0.01 ).Compared with group I/R,no significant change was found in the parameters mentioned above in group Spo+ L (P ＞ 0.05),and the levels of CK-MB,LDH and cTnI and IS/AAR ratio were significantly decreased,and the ratios of p-Akt/Akt and p-eNOS/eNOS were significantly increased in groups Spo and Spo + D ( P ＜ 0.05 or 0.01 ).There was no significant difference in the parameters mentioned above between group Spo and group Spo + D (P ＞ 0.05).Conclusion PI3K-Akt-eNOS signaling pathway mediates the attenuation of myocardial I/R injury by sevoflurane postconditioning in rats.

Objective Toobservetheindication, safetyandefficacyofanew immunosuppressant Rituximab in kidney transplantation. MethodsFive patients, who were diagnosed as antibody mediated rejection (AMR) from December 2010 to June 2011, were treated with single dose of Rituximab (500 mg) and followed up for 6 months. The clinical data, such as age, gender, onset of illness, induction therapy, maintaining therapy, allograft function, change of PRA, opportunistic infection and other complications were collected and retrospectively analyzed to evaluate the safety and efficacy of Rituximab used in AMR patients. ResultsAfter Rituximab therapy, all the patients had improved renal function measured by sera creatinine level: 4 cases retumed to normal, and 1 keep stable. Series of allograft biopsy demonstrated obviously reduced C4d deposition in nephridial tissue after treatment. One patient developed CMV viremia, another had urinary infection, but no one had lifethreatening infection during the follow-up period. The survival rate of human and allograft was both 100 ％. Conclusion Rituximab has a good efficacy and safety in treatment of AMR after renal transplantation.

Objective To determine the feasibility of using a special auditory intervention to detect intraoperative awareness in children under general anesthesia. Methods Thirty-four ASA Ⅰ or Ⅱ children aged 5-15 yr weighing 16-73 kg undergoing scoliosis were enrolled in this study.Intraoperative wake-up test was performed during operation.Two easily identified animal noises(60 dB,lasting 15 s)were played through head phones before induction of anesthesia and immediately after the intraoperative wake-up test. Children were interviewed on the 1st and 5th-7th days after surgery to assess their awareness of intraoperative wake-up test and special auditory intervention. Results Four children were excluded from the study because of restlessness during the intraoperative wake-up test or their refusal to be interviewed. From the remaining 30 children,4 children were suspected to be aware of intraooperative wake-up test. Awareness was comfirmed in 3 children and suspected in one child.The incidence of awareness of intraoperative wake-up test was 10%.But only one of them could tell the animal sound played during the wake-up test.All the patients in this study had explicit recall of the animal noises played before induction of anesthesia.Conclusion Special auditory intervention can not detect intraoperative awareness in children unnder general anesthesia.

Objective To evaluate the efficacy of percutaneous transluminal angioplasty (PTA) in the treatment of transplanted renal artery stenosis (TRAS)-induced renal dysfunction and hypertension. Methods Between July 1998 and January 2007, PTA was performed on 16 patients with RTAS. Color Doppler uhrasonography preceded the intra-arterial angiographic investigation,with false-negative results in 18. 75 % of patients. Sixteen cases of TRAS were examined at 1 st week,6th month and 13 years after PTA. Hypertension improvement was defined as mean arterial pressure decrease of at least 15 % from the pre-PTA value. Graft function was evaluated by SCr levels, and the improvement was defined as a 20% change. Results Angioplasty was technically feasible in 100 %.Sixteen patients with RTAS were cured clinically. During the follow-up period, graft function was improved in 81.25 %, 68. 75 %, 62. 5 %, 56. 25 %, 50 % of patients respectively at 1st week, 6th month and 1-3 years after PTA. The blood pressure was decreased in 62. 5%, 75 %, 75 %,56. 25 %, 50 % of patients respectively, but no patient remained hypotensor medication free.Conclusion PTA improved renal dysfunction and hypertension induced by TRAS, and it is a safe and effective treatment for TRAS.