Cryoablation therapy with explicit anti-tumor mechanisms and histopathological manifestations has a long history.A large number of clinical practice has shown that cryoablation therapy is safe and effective,making it an ideal tumor treatment method in theory.Previously,its efficacy and clinical application were constrained by the limitations of refrigerants and refrigeration equipment.With the development of the new generation of cryoablation equipment represented by argon helium knives,significant progress has been made in refrigeration efficien-cy,ablation range,and precise temperature measurement,greatly promoting the progression of tumor cryoablation technology.This consensus systematically summarizes the mechanism of cryoablation technology,indications for oral mucosal melanoma(OMM)cryotherapy,clinical treatment process,adverse reactions and management,cryotherapy combination therapy,etc.,aiming to provide reference for carrying out the standardized cryoablation therapy of OMM.

OBJECTIVE To investigate the effect of gene CD47 on immune microenvironment of lung squamous cell carcinoma and predict its immune response. METHODS The differences in the expression of CD47 between lung squamous cell carcinoma tissues and normal tissues were compared through The Cancer Genome Atlas(TCGA) databases. CD47 involved immune-related pathways were analyzed by Gene Set Enrichment Analysis(GSEA). Tumor Immune Estimation Resource(TIMER) databases and TISIDB databases(http://cis.hku.hk/TISIDB/TISIDB) were employed to analyze the relationship between CD47 gene expression in lung squamous cell carcinoma and immune cells infiltration in the tumor microenvironment. And further explored the associations between CD47 and other immune-related molecules such as PD-1, PD-L1, CTLA-4, IDO-1. Using the pRRophetic and TIDE algorithms, this study predicted the correlation between the expression of CD47 gene and chemotherapy drug sensitivity as well as the response to immunotherapy. RESULTS Compared with normal tissues, CD47 was lower expressed in lung squamous cell carcinoma tissues. It was found that CD47 could activate immune-related pathways such as the inflammatory response, interferon alpha response, interferon gamma response and JAK-STAT pathway. In lung squamous cell carcinoma tissues, the expression level of CD47 was significantly positively correlated with the infiltration level of B-cells, CD8T-cells, CD4T-cells, macrophage, neutrophils and dendritic cells according to the two databases. At the same time, CD47 showed a positive correlation with other immune checkpoints such as PD-1, PD-L1, CTLA-4, IDO1. The result of chemotherapy sensitivity indicated that in lung squamous cell carcinoma tissues, IC50 values of high expression CD47 group were lower than that for chemotherapy drugs such as sunitinib, dasatinib, doxorubicin and gemcitabine. However, the response rate to immunotherapy was lower in the low expression group for the immunotherapy. CONCLUSION CD47 is low expressed in lung squamous tissue, it can activate immune related pathways, and has a high response rate to immunotherapy. It can be used as a novel molecular marker for lung squamous cell carcinoma.

Objective:To explore the epidemiological characteristics and clinical outcomes of patients diagnosed with novel coronavirus (COVID-19) pneumonia (NCP), and to provide clinical reference for subsequent epidemic prevention and control.Methods:The pathogenesis, clinical characteristics and clinical outcomes of 62 patients with imported novel coronavirus COVID-19 pneumonia in Qingdao area from January 21, 2020 to April 6, 2020 were analyzed.Results:There were 24 males and 38 females, aged from 5 to 91 (52.5±15.7)years. Among them, 31 cases (31/62, 50%) were family cluster disease, 11 cases (17.74%) were nosocomial cross infection, 14 cases (22.58%) came back from Wuhan to Qingdao and community contact infection, and 6 cases (9.68%) were imported from abroad. The longest time from fever to diagnosis was 26 days. Clinical classification: 40 cases were mild, 13 were general, 7 were severe, 2 were critically ill. 61 cases were discharged from hospital, and 1 was dead. The clinical cure rate was 98.39%. The average stay was (18.5±4.8)days. All patients were treated combined with Chinese medicine on the basis of symptomatic treatment.Conclusions:The clinical symptoms of imported NCP are relatively mild, and mild cases were more, the recovery rate was higher. The intervention strategy of traditional Chinese medicine is conducive to rehabilitation. The external input cases and symptomless infected patients are important for epidemic situation in the future.

Objective:To investigate the correlation between RNF213 gene p. R4810K polymorphism and posterior cerebral artery involvement in Chinese children with familial moyamoya disease.Methods:Children with familial moyamoya disease admitted to the Department of Neurosurgery, the Fifth Medical Center of PLA General Hospital from August 2004 to June 2018 were enrolled, and they were divided into posterior cerebral artery involved group and posterior cerebral artery uninvolved group. RNF213 gene p. R4810K single nucleotide polymorphism was detected. Multivariate logistic regression analysis was used to determine the independent risk factors for posterior cerebral artery involvement. Results:A total of 65 children with familial moyamoya disease were enrolled. Their age was 6.98±4.46 years and 37 (56.9%) were male. The first symptom of 55 children (84.6%) was cerebral ischemia, and 37 (56.9%) involved posterior cerebral artery. There were 3 (4.6%) children with p. R4810K AA genotype, 26 (40.0%) with GA genotype, and 36 (55.4%) with GG genotype. The p. R4810K genotype distribution in the posterior cerebral artery involved group was statistically different from that in the uninvolved group (GA+ AA genotype: 56.8% vs. 28.6%; χ2=5.124, P=0.024), and there were no statistical difference in gender, age, first symptom, and genetic pattern. Multivariate logistic regression analysis showed that after adjusting the first onset age and gender, p. R4810K G>A mutation was the only independent risk factor for posterior cerebral artery involvement (odds ratio 3.240, 95% confidence interval 1.082-9.705; P=0.020). Conclusion:The p. R4810K polymorphism of RNF213 gene is associated with posterior cerebral artery involvement in Chinese children with familial moyamoya disease.

Objectives To evaluate the effectiveness and safety of peramivir trihydrate in patients with influenza.Methods This was a randomized,double-blind,double-dummy,placebo and positive control,multicenter clinical trial,comparing peramivir trihydrate with oseltamivir and placebo.The inclusive criteria were 15-70 years old,onset within 48 h,positive rapid influenza antigen test,and febrile(＞38℃) accompanied with at least two associated symptoms.The severe cases complicated with chronic pulmonary and cardiac diseases,malignancies,organ transplantation,hemodialysis,uncontrolled diabetes,immunocompromised status,pregnancy and coexistence of bacterium infections were excluded.All patients were randomized 2:2:1 to receive peramivir,oseltamivir and placebo respectively.The primary endpoint was the disease duration,the secondary endpoints included time to normal axillary temperature and normal living activities,viral response,and adverse effects.Results Following informed consent,133 patients were included in this study.Four patients were exclude due to missing medical records,not fitting inclusion or exclusion criteria and poor compliance.A total of 129 patients were finally analyzed,including 49 cases,54 cases and 26 cases in peramivir group,oseltamivir group and placebo group.The median disease duration were 96 (76,120)hours,105(90,124) hours,and 124 (104,172)hours in three groups respectively(P＞0.05).The time to normal axillary temperature,normal living activities and viral response were not significantly different in three groups(P＞0.05).Conclusion The value of antiviral therapy in patients with mild influenza needs to be further determined.

The accelerating growth of the public microbial genomic data imposes substantial bur-den on the research community that uses such resources. Building databases for non-redundant ref-erence sequences from massive microbial genomic data based on clustering analysis is essential. However, existing clustering algorithms perform poorly on long genomic sequences. In this article, we present Gclust, a parallel program for clustering complete or draft genomic sequences, where clustering is accelerated with a novel parallelization strategy and a fast sequence comparison algo-rithm using sparse suffix arrays (SSAs). Moreover, genome identity measures between two sequences are calculated based on their maximal exact matches (MEMs). In this paper, we demon-strate the high speed and clustering quality of Gclust by examining four genome sequence datasets. Gclust is freely available for non-commercial use at https://github.com/niu-lab/gclust. We also introduce a web server for clustering user-uploaded genomes at http://niulab.scgrid.cn/gclust.

Objective To preliminarily explore the clinical features,treatment,and outcomes of moyamoya disease in the elderly.Methods The clinical data of the elderly patients with moyamoya disease (aged ＞ 60 years) admitted to the Department of Neurosurgery,the 307th Hospital of PLA from May 2007 to July 2016 were collected retrospectively.Their clinical features,imaging features,and surgical outcomes were analyzed.Results A total of 68 patients were enrolled,including 35 females (51.47％) and 33 males (48.53％).The ratio of male to female was 1:1.06.The age at the time of diagnosis of moyamoya disease was 62.82 ±3.08 years.Fifty-two patients (76.5％) had vascular risk factors.The most common clinical manifestation was cerebral ischemia (n =61,89.7％).Thirty of them (44.1％) presented as transient ischemic attack.The Suzuki staging of most patients was 4-6 (71.6％),12 patients (17.6％) complicated with posterior cerebral artery stenosis or occlusion.Thirty-one patients were treated with encephalo-duroarterio-synangiosis (EDAS).Among them,17 patients underwent bilateral surgery and 14 underwent unilateral surgery.The incidence of perioperative infarction or hemorrhage was 5.6％ (2 patients developed cerebral infarction and 1 patient developed cerebral hemorrhage);37 patients received conservative treatment.During the follow-up period,5 patients developed cerebral infarction (1 in the surgical treatment group and 4 in the conservative treatment group);there was no significant difference between the 2 groups.There were no significant differences in age,sex,vascular risk factor,clinical symptoms,and preoperative modified Rankin Scale (mRS) scores between the 2 groups.Cerebral angiography was performed 6-9 months after operation in the surgical treatment group.A total of 24 cerebral hemispheres were evaluated by Matsushima typing,of which 17 (70.8％) were excellent.During the follow-up period,the proportion of patients with clinical outcome excellent (the mRS score was 0) (Z =-5.268,P ＜ 0.00l) and clinical improvement (the mRS score was improved ≥ 1 compared to the baseline) (Z =-3.780,P ＜ 0.001) were significantly higher than the conservative treatment group.Conclusions The clinical symptoms of old patients with moyamoya disease were mainly cerebral ischemia.Most of them had vascular risk factors,and the imaging manifestations showed higher Suzuki staging.The perioperative risk of EDAS in the old patients with moyamoya disease was lower.It might be an effective method to prevent clinical symptoms progress and improve the outcomes.

Objective:To observe the effects of isobavachalcone (IBC)on the proliferation and apoptosis of tongue squamous cell carcinoma Tca-8113 cells,and to explore their mechanisms. Methods:The Tca8113 cells cultured in vitro were divided into control group and different doses (0, 10, 20,40, 80 μmol · L-1 )of IBC groups.The inhibitory rates of cell proliferation were detected by MTT method.The apoptosis was detected by flow cytometry.Western blotting method was used to detect the expressions of Akt,p-Akt,Erk,p-Erk,Bax, Bcl-2 and Caspase-3 proteins in the Tca8113 cells in various groups.Results:The MTT results showed that the inhibitory rates of proliferation of Tca8113 cells were increased in a concentration-and time-dependent manner;the IC50 at 12,24 and 48 h were (285.13±8.97), (132.40±7.76),and (58.56±5.93)μmol·L-1 ,respectively;and there were significant differences between different time points (P <0.05).The FCM results showed that the apoptotic rates of Tca8113 cells in 20 and 40 μmol·L-1 IBC groups at 48 h were (8.21 ±2.32)% and (22.45 ± 1.18)%, respectively; compared with control group (1.69% ± 0.65%), the differences were statistically significant (P <0.05).The Western blotting results showed that the expression levels of Bcl-2,p-Akt,and p-Erk, in 20 and 40 μmol·L-1 IBC groups were decreased and the expression levels of Bax were increased compared with control group (P < 0.05);the expression levels of Akt and Erk had no significant changes (P > 0.05 ).The expression levels of Caspase-3 in Tca8113 cells in 40 μmol·L-1 IBC group at 48 h were increased compared with control group (P <0.05).Conclusion:IBC could inhibit the proliferation and induce the apoptosis of Tca8113 cells;Akt and Erk signaling pathway may be the pathway of IBC to induce the apoptosis of tumor cells.

Objective To summarize the clinical experience of surgical intervention for cardiac neoplasm in a fetus . Methods A 32-year-old pregnant woman was admitted to our hospital for complaint of fetal cardiac neoplasm .A separated het-erogenic cardiac occupying lesion was identigied at right atrium of the fetus by echocardiography , whose size is 2.85 cm ×2.25 cm, but the pathogenic origin still remained uncertain, maybe originate from ether pericardium or atrium.The annulus of tri-cuspid valve was compressed nearly 50% with the presence of amount of pericardial effusion.The fetal heart rate decreased at some fetal position resulting in the compression to the heart.So an Ex-utero Intrapartum Therapy(EXIT) procedure was per-formed under the supply of placenta at the 32 weeks of pregnancy.Cesarean section was performed with intact umbilicus and fe-tal circulation by obstetricians.Consequently, the median sternotomy of this fetus and pericardiotomy were performed , with 30 ml clear pericardial effusion drained .The tumor was confirmed to be giant right atrial neoplasm after the intraoperative explora-tion.Considering on the high risk of the cardiopulmonary bypass and limited time for EXIT , the giant atrial neoplasm was left alone with delayed sternum closure after the effectively decompression of the heart .The omphalotomy was successfully per-formed after the EXIT surgery.The neoplasm resection and the repair for its defect on right atrium were performed with cardiop-ulmonary bypass 2 days later.Results Convalesce of this mother was quite good after cesarean resetion .Hemodynamics of the premature baby was satisfatory after the resection of right atrial neoplasm which pathological report was benign hemangioma . Conclusion Via multiple disciplines collaboration , EXIT intervention for fetus is feasible and safe under adequate prepara-tion.

Objective To investigate the clinical features of subarachnoid hemorrhagic moyamoya disease and the therapeutic effect of encephalo-duro-arterio-synangiosis (EDAS). Methods The clinical and imaging data of 38 patients with subarachnoid hemorrhagic moyamoya disease admitted to the Department of Neurosurgery,the 307th Hospital of PLA from January 2002 to April 2013 were analyzed retrospectively. Thirty-five patients underwent unilateral or bilateral EDAS (64-sides underwent EDAS,4 patients with aneurysms underwent endovascular embolization first),and 3 patients did not undergo any surgery. Results (1)Subarachnoid hemorrhagic moyamoya disease accounted for 10. 8%(38/353)of all the hemorrhagic moyamoya disease admitted in hospital over the same period,including 37 adults and 1 child. The male to female ratio was 1∶3. 22 (9/29),and the age of onset was 12 to 59 years. The mean age of patients was 39 ± 11 years. Four patients were combined with aneurysms. There were no significant differences in the distribution of Suzuki stage,anterior choroidal artery dilatation and posterior communicating artery dilatation in the remaining 34 patients without aneurysms between the bleeding sides and non-bleeding sides (P>0.05). (2 ) The patients were followed up for 13 -125 months (mean 51 ± 27 months ),two patients had rebleeding,one of them was intraventricular hemorrhage,the other was parenchymal hemorrhage. The postoperative modified Rankin score (mRS)was significantly lower in 35 patients whom were treated with EDAS. Compared with before surgery,there was significant difference (P<0. 05). The re-examination of positron emission tomography (PET)for 16 patients at 3 to 19 months after surgery showed that among the 23 surgically treated hemispheres,the cerebral metabolisms of 17 hemispheres were improved after surgery, and 6 did not have any change after surgery. The re-examination of whole brain digital subtraction angiography (DSA)at 5 to 30 months after surgery in 13 patients showed that revascularizations in 19 of 23 surgical hemispheres were effective. Conclusion Subarachnoid hemorrhagic moyamoya disease often occurs in adults,and women are more common. EDAS can achieve good revascularization effect and improve brain metabolism of patients,and thus relieve the symptoms of cerebral ischemia.