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.

Cerebral small vessel disease（CSVD）is one of the most major causes of cognitive dysfunction. With the acceleration of population aging in China and the development of medical imaging technology，the incidence rate of CSVD has been on the rise，and related cognitive dysfunction has become an increasingly concerning issue. Due to its insidious onset，slow progression，and lack of early warning signs，CSVD is typically found when moderate or severe cognitive impairment or even dementia already occurs，often causing irreversible damage and heavy medical burden to patients. This article reviews the impact of different imaging types of CSVD and total MRI load on cognitive function to further understand the relationship between CSVD and cognitive function，aiming to provide a reference for the identification and prevention of cognitive dysfunction caused by CSVD.

Objective To establish a fluorescent assay for rapid detection of Plasmodium falciparum based on recombinaseaided amplification (RAA) and CRISPR-Cas12a system,and to preliminarily evaluate the diagnostic efficiency of this system.. Methods The 18S ribosomal RNA (rRNA) gene of P. falciparum was selected as the target sequence, and three pairs of RAA primers and CRISPR-derived RNA (crRNA) were designed and synthesized. The optimal combination of RAA primers and crRNA was screened and the reaction conditions of the system were optimized to create a fluorescent RAA/CRISPR-Cas12a system. The plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 was generated, and diluted into concentrations of 1 000, 100, 10, 1 copy/μL for the fluorescent RAA/CRISPR-Cas12a assay, and its sensitivity was evaluated. The genomic DNA from P. vivax, P. malariae, P. ovum, hepatitis B virus, human immunodeficiency virus and Treponema pallidum was employed as templates for the fluorescent RAA/CRISPR-Cas12a assay, and its specificity was evaluated. Fifty malaria clinical samples were subjected to the fluorescent RAA/CRISPR-Cas12a assay and nested PCR assay, and the consistency between two assays was compared. In addition, P. falciparum strain 3D7 was cultured in vitro. Then, the culture was diluted into blood samples with parasite densities of 1 000, 500, 200, 50, 10 parasites/μL with healthy volunteers’ O-positive red blood cells for the RAA/CRISPR-Cas12a assay, and the detection efficiency was tested. Results The Pf-F3/Pf-R3/crRNA2 combination, 2.5 μL as the addition amount of B buffer, 40 min as the RAA reaction time, 37 °C as the reaction temperature of the CRISPR-Cas12a system were employed to establish the fluorescent RAA/CRISPR-Cas12a system. Such a system was effective to detect the plasmid containing 18S rRNA gene of the P. falciparum strain 3D7 at a concentration of 1 copy/μL, and presented fluorescent signals for detection of P. falciparum, but failed to detect P. ovum, P. malariae, P. vivax, T. pallidum, hepatitis B virus or human immunodeficiency virus. The fluorescent RAA/CRISPR-Cas12a system and nested PCR assay showed completely consistent results for detection of 50 malaria clinical samples (kappa = 1.0, P < 0.001). Following 6-day in vitro culture of the P. falciparum strain 3D7, 10 mL cultures were generated and the fluorescent RAA/CRISPR-Cas12a system showed the minimal detection limit of 50 parasites/μL. Conclusion The fluorescent RAA/CRISPR-Cas12a system is rapid, sensitive and specific for detection of P. falciparum, which shows promising value for rapid detection and risk monitoring of P. falciparum.

ObjectiveTo develop a new mosquito trap and evaluate its effectiveness in Aedes albopictus monitoring in an urban community. MethodsThe first-round field test was conducted in a well-greened residential neighborhood with high Aedes albopictus density calculated by human landing catch in August 2022， in Minhang District， Shanghai. 65 new test containers with different designs were randomly deployed in the field. These trap containers were designed by different material， color， hole size， shape and top cover colors. The results of mosquito and its egg trapping were observed and recorded continuously for 7 days. In September 2022， the second round of experiment was conducted. After comparison， an optimized trap container was designed. 140 new trap containers were compared with 140 traditional ones to evaluate the effectiveness. SPSS 22.5 software was used for statistical analysis. ResultsThere were significant changes in the outcome of mosquito and its egg trapping on the 4th and 7th day of the pre-experiment deployment of the three kinds of containers. The 200 ml container cup with three round holes in the body had the best effect on trapping mosquitos and eggs， with a positive rate of 45.5% （5/11）， while other type of contrainer cup only had a positive rate of 12% （6/50） in trapping mosquitos and eggs. Statistical significant differences （χ²=45.443，P <0.001） existed between the two tpyes of cups within the first kind of trapping container. The second kind of trapping container showed round and oval holes had better effect on mosquito trapping， with a positive rate of 55.6% （5/9）. The third kind of hanging container can resist strong wind. The best mosquito trapping effect of three round holes was 66.7% （2/3）. The container wall should not be made of rough material， which was easy to cause the escape of adult mosquitoes. Combined with the three types of containers， the new contariner should be top transparent， and the top 1/3 of the side should paint black with 3 round holes. The effective recovery rate of the new container was higher than that of the traditional one under the exclusion of human factors， and had the advantage of preventing rain， wind and rollover， but the effect of trapping mosquito and its egg was comparable to the traditional one（χ²_D4=0.197，P=0.658； χ²_D7=0.125 ，P=0.724）. ConclusionThis new type of mosquito trap is worth further exploration and research， which can improve the trapping efficiency， overall recovery rate and the accurate confirmation of the mosquito density.

In order to improve the yield and simplify the operation, the synthesizing process of JAK3 inhibitor tofacitinib citrate was improved based on the analysis of the methods previously published.Using 2, 4-dichloro-7H-pyrrolo [2, 3-d] pyrimidine and (3R, 4R)-1-benzyl-N, 4-dimethylpiperidin-3-amine dihydrochloride as starting materials, tofacitinib citrate was obtained through four steps of nucleophilic substitution, catalytic transfer hydrogenation, cyanide acetylation and citrate salt, and its crystal form was consistent with the original research.After optimization, the yield was better than those reported in literature, and the mild reaction conditions were suitable for industrial production.

Objective:To analyze the changes of subfoveal choroidal thickness (SFCT) and choroidal volume in type 2 diabetes patients with diabetic retinopathy (DR) through automated choroid segmentation.Methods:A cross-sectional study was conducted.Sixty-four patients (88 eyes) with type 2 diabetes diagnosed in The First Affiliated Hospital with Nanjing Medical University from May 2016 to May 2017 were enrolled, including 27 males (32 eyes) and 37 females (56 eyes), with an average age of (62.7±7.4) years and an average diabetes duration of (13.7±7.2) years.Best corrected visual acuity, slit lamp ophthalmoscopy, indirect ophthalmoscopy and spectral domain optical coherence tomography (SD-OCT) examination were carried out among all the patients.The patients were divided into non-DR (NDR) group (28 eyes), nonproliferative DR (NPDR) group (36 eyes) and proliferative DR (PDR) group (24 eyes) according to the Early Treatment Diabetic Retinopathy Study.Nineteen age-matched normal subjects (32 eyes) were enrolled as normal control group.SFCT and choroidal volume were measured through automated choroid segmentation based on SD-OCT with enhanced depth imaging.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital with Nanjing Medical University (No.2017-SR-176).Results:Mean SFCT and choroidal volume of normal control group, NDR group, NPDR group and PDR group were (277.8±45.3)μm and (8.8±1.0)mm 3, (220.9±38.7)μm and (7.6±1.3)mm 3, (231.8±26.7)μm and (8.2±0.9)mm 3, (252.0±42.2)μm and (8.7±1.0)mm 3, respectively, with significant overall differences among the groups ( F=14.615, P<0.001; F=7.711, P<0.001). Mean SFCT and choroidal volume were significantly smaller in NDR and NPDR group than those in normal control group (both at P<0.05). Mean SFCT and choroidal volume of PDR group was greater than those of NDR group (both at P<0.05). Conclusions:SFCT and choroidal volume decrease during the early course of diabetes and increase significantly as DR worsens from NDR to PDR.

Objective:To explore how to optimize selection, acquisition, and matching for the use of an aged donor liver.Methods:A case of 81-year-old senior donor liver transplantation performed at our institution is reported, the donor was an 81-year-old female who was brain-dead and the recipient was a 21-year-old male who underwent liver transplantation because of chronic hepatitis B complicated with hepatocellular carcinoma. Combined with the literature, the preliminary experience of age-related marginal donor liver in clinical liver transplantation was discussed.Results:The recipient recovered smoothly. Aspartate aminotransferase(AST)and glutamate-pyruvate transaminase(ALT)were 1131 U/L and 846 U/L on the first postoperative day, and decreased gradually to normal until 2 weeks later.Total bilirubin(TBIL)56.4umol/L on the first day and decreased to 22.1umol/L on the third day after operation, however, it began to rise on postoperative day 5 to a maximum of 66.0 umol/L, which decreased to normal about 2 months after surgery. During the follow-up, AST, ALT, TBIL and other indexes were normal, and AFP decreased progressively.Conclusions:Using of elderly liver donors may achieve satisfactory results, asifthe elderly donors been evaluatedcarefullyandthe low-risk recipients been selected.

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 summarize our experience in 13 cases of intestinal autotransplantation (IATx) after resection of lesions involving the roots of mesenteric vessels.Methods:The clinical data of 13 patients undergoing IATx in Xijing hospital were retrospectively analyzed. The etiology, surgical procedure and complications were analyzed. The patients were followed up by telephone and regular evaluations.Results:All 13 cases of IATx were successfully completed. For 12 patients who were diagnosed with tumors involving the mesenteric roots, the tumors were removed for cure intent avoiding massive intestinal resection. Pancreaticoduodenectomy was carried out simultaneously in 11 cases. The postoperative complication rate was 85% (11/13). The autograft was resected in 1 patient on the 1st postoperative day due to necrosis from mesenteric thrombosis, leading to short bowel syndrome. One-year survival was 69% (9/13). Among 4 deaths, 1 patient died of liver metastasis, and another died of liver and lung metastases. Five patients were alive 2 years postoperatively.Conclusion:IATx while-technically challenging, avoiding short small bowel syndrome in properly selected patients after resection of lesions especially benign ones involving the roots of mesenteric vessels that were traditionally considered to be "unresectable".

Objective: To investigate the immunohistochemical staining of anaplastic lymphoma kinase (ALK; clone 1A4) in pediatric medulloblastoma (MB). Methods: Molecular subtyping was performed based on the NanoString and sequencing techniques for 44 pediatric MB cases at Children′s Hospital, Zhejiang University School of Medicine from 2014 to 2017. ALK expression was detected with EnVision immunhistochemistry using ALK clone 1A4 on whole section. Statistical analyses were performed to evaluate the correlation of protein expression with molecular subgroups. Results: The age ranged from 0.5 to 13.0 years with an average age of 5.8 years. There were 28 males and 16 females, and 31 classic, 5 desmoplastic nodular, 3 extensive nodular and 5 large cell/anaplastic MBs. Except three cases was unable classified, 41 MBs were classified into the four molecular groups: 5 in WNT group, 12 in SHH group, 9 in Group 3 and 15 in Group 4. Thirteen of 44 MB cases were positive staining for ALK, and the positive rate was 29.5%. Six cases were strong reaction, and 7 cases were weak. The expression of ALK at the protein level was associated with the WNT group (P<0.01). The characteristic perinuclear dot-like staining was only showed in WNT group. Conclusions The ALK immunhistochemistry using antibody clone 1A4 is a useful marker for the molecular subgroup detection of MB. The strong staining and perinuclear dot-like staining indicate as WNT group.