Objective·To establish a mouse model with dormant cancer and no obvious metastasis by combining the preimmune strategy with the mVenus-p27K-cell G0 phase indicator system,the DTR-HSV/TK suicide gene system,and the Luc2-tdTomato tracer system.Methods·The KPC1199 mouse pancreatic cancer cell line was transfected with the mVenus-p27K-cell G0 phase indicator system,the DTR-HSV/TK suicide gene system,and the Luc2-tdTomato tracer system to construct a stable expression cell line,KPC1199-PDL.After being cultured in the serum-free condition,KPC1199-PDL cells were sorted into mVenus(+)cells and mVenus(-)cells by flow cytometry,and the expression of G0 phase-related genes was verified by real-time fluorescence quantitative PCR(qPCR).Sensitivity of KPC1199-PDL cells to diphtheria toxin(DTX)and ganciclovir(GCV)was evaluated by CCK-8 assay.A transsplenic portal vein-hepatic metastasis model was constructed in wild-type C57BL/6 mice to validate the function of KPC1199-PDL cells in vivo by immunofluorescence technology.The KPC1199-PDL cells were injected subcutaneously into C57BL/6 mice,followed by in situ injection of DTX and GCV to ablate subcutaneous tumors 5 d later,to obtain preimmunized mice.The transsplenic portal vein-hepatic metastasis models were constructed in these mice.Bioluminescence imaging was used to evaluate subcutaneous tumor ablation and hepatic metastasis in the mice,and immunofluorescence assay was used to detect the distribution and dormant state of tumor cells in the livers of preimmunize mice.Results·The three tool systems were stably expressed in KPC1199-PDL cells,and their proliferative ability was not affected.In the serum starving condition,some KPC1199-PDL cells expressed the mVenus protein,indicating entry into the G0 phase;the mVenus(+)cells sorted out by flow cytometry exhibited significantly higher expression of G0 phase-related genes(all P<0.05)and significantly lower expression of the proliferation-related gene compared with mVenus(-)cells(P<0.05).The CCK-8 assay demonstrated high sensitivity of KPC1199-PDL cells to DTX and GCV.In vivo experiments confirmed that KPC1199-PDL cells could be effectively traced through tdTomato protein expression,and could indicate entry into the G0 phase through mVenus protein expression.Following subcutaneous tumor implantation and drug ablation,preimmunized mice were successfully obtained.In the subsequent transsplenic portal vein-hepatic metastasis model,no metastatic signals were observed in the liver by bioluminescence imaging,but single or small clusters of G0 phase tumor cells expressing both mVenus and tdTomato,not expressing the proliferation marker Ki67,were detected in liver tissue sections by immunofluorescence analysis.Conclusions·A recognizable and traceable dormant cancer model is constructed with the combination of the preimmune mouse model of pancreatic cancer,the mVeneus-p27K-indicator system,the DTR-HSV/TK suicide gene system,and the Luc2-tdTomato tracer system.

Objective:To observe the clinical phenotype of patients with CACNA1F gene variant. Methods:A retrospective clinical study. From January 1, 2022 to October 1, 2023, 36 patients with CACNA1F gene mutation-related eye diseases diagnosed by clinical examination and genetic testing in Changsha Aier Eye Hospital and Jinan Purui Eye Hospital were included in the study. All patients underwent best-corrected visual acuity (BCVA), medical optometry, fundus color photography, optical coherence tomography, full-field electroretinography (ERG), nystagmus examination, and genetic whole-exon sequencing. BCVA was performed using log visual acuity charts and converted to (logMAR). The nystagmus examination was performed using a helmet-mounted multifunctional video eye movement recording system. The clinical phenotypic characteristics were observed. Results:At total of 36 patients were male, aged was (6.69±5.26) years. There were 36 cases of myopia (38.89%, 14/36), and the spherical equivalent was (?3.01±4.84) D. There were 14 different genetic variants including 7 cases of pathogenic variants, 20 cases of suspected pathogenic variants and 9 cases of unknown pathogenic variants, respectively. logMAR BCVA was 0.67±0.27; 26 patients had optic nerve atrophy (72.22%, 26/36); 6 had optical nerve hypoplasia (16.67%, 6/36). Fundus pigment dysplasia with mild iris transillumination was found in 4 cases (11.11%, 4/36). There were 5 cases of foveal dysplasia (Thomas grade) 1 (13.89%, 5/36). In full-field ERG examination, the B-wave reduction of the maximum mixed reaction of dark adaptation showed a negative waveform, and the amplitude of the shock potential was seriously reduced. The main phenotypes were residual type (residual dark adaptation 0.01 reaction wave and bright adaptation 3.0 reaction wave, and the response decreased at 30 Hz to a double-peak wide wave), dominant type of bright adaptation decreased (all light adaptation extinguished, all dark adaptation extinguished), and total extinction type (all light adaptation extinguished). Among them, 10 cases presented with residual ERG (27.78%, 10/36), 8 cases with photopic reduced ERG (22.22%, 8/36) and 18 with extinguished ERG (50.00%, 18/36). Low amplitude and high frequency pendulum (PLAHF) nystagmus waverforms were found in 32 cases (88.89%, 32/36), head oscillation in 27 cases (75.00%, 27/36) and chin up abnormal head posture in 26 cases (72.22%, 26/36), respectively.Conclusions:CACNA1F Gene variant eye diseases had diverse clinical phenotype. Clinical phenotype of PLAHF nystagmus is closely related with CACNA1F gene variant eye disease.

Objective In tongue diagnosis, the location, color, and distribution of spots can be used to speculate on the viscera and severity of the heat evil. This work focuses on the image analysis method of artificial intelligence (AI) to study the spotted tongue recognition of traditional Chinese medicine (TCM). Methods A model of spotted tongue recognition and extraction is designed, which is based on the principle of image deep learning and instance segmentation. This model includes multiscale feature map generation, region proposal searching, and target region recognition. Firstly, deep convolution network is used to build multiscale low- and high-abstraction feature maps after which, target candidate box generation algorithm and selection strategy are used to select high-quality target candidate regions. Finally, classification network is used for classifying target regions and calculating target region pixels. As a result, the region segmentation of spotted tongue is obtained. Under non-standard illumination conditions, various tongue images were taken by mobile phones, and experiments were conducted. Results The spotted tongue recognition achieved an area under curve (AUC) of 92.40%, an accuracy of 84.30% with a sensitivity of 88.20%, a specificity of 94.19%, a recall of 88.20%, a regional pixel accuracy index pixel accuracy (PA) of 73.00%, a mean pixel accuracy (mPA) of 73.00%, an intersection over union (IoU) of 60.00%, and a mean intersection over union (mIoU) of 56.00%. Conclusion The results of the study verify that the model is suitable for the application of the TCM tongue diagnosis system. Spotted tongue recognition via multiscale convolutional neural network (CNN) would help to improve spot classification and the accurate extraction of pixels of spot area as well as provide a practical method for intelligent tongue diagnosis of TCM.

Objective: The current study aims to find out the potential reasons why most schizophrenia patients have a relatively low sensitivity to the classification of emotional facial expressions. Methods: By using an emotional categorical perception task, eighty-three schizophrenia patients and seventy-one healthy adults are provided with morphed emotional continuums with two emotional facial expressions (a positive emotional valence: happy; a negative emotional valence: sad). Results: Through comparing the difference between schizophrenia patients and healthy adults in the processes of estimating facial expressions with ambiguous emotions, we find that the pattern of emotional categorical perception for schizophrenia patients is significantly different from that of healthy controls when they process signals on the local facial areas. Compared to healthy people, schizophrenia patients have a significantly separate classification pattern in processing emotional signals between the eyes and mouth regions. It indicates that compared to healthy adults, schizophrenia patients have larger conflicts in integrating emotional signals from different facial areas. To overcome conflicts, more cognitive resources are required. Unfortunately, the lack of cognitive resources leads to the failure of integration, which further increases the difficulty of estimating facial expressions with ambiguous emotions, and finally leads to the relatively low sensitivity of emotional facial expressions classification. Conclusion To sum up, the deficit of abnormal perceptions of emotional facial expressions in schizophrenia patients results from an integrated deficit of signals on facial areas.

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has occasioned worldwide alarm. Globally, the number of reported confirmed cases has exceeded 84.3 million as of this writing (January 2, 2021). Since there are no targeted therapies for COVID-19, the current focus is the repurposing of drugs approved for other uses. In some clinical trials, antiviral drugs such as remdesivir (Grein et al., 2020), lopinavir/ritonavir (LPV/r) (Cao et al., 2020), chloroquine (Gao et al., 2020), hydroxychloroquine (Gautret et al., 2020), arbidol (Wang et al., 2020), and favipiravir (Cai et al., 2020b) have shown efficacy in COVID-19 patients. LPV/r combined with arbidol, which is the basic regimen in some regional hospitals in China including Zhejiiang Province, has shown antiviral effects in COVID-19 patients (Guo et al., 2020; Xu et al., 2020). A retrospective cohort study also reported that this combination therapy showed better efficacy than LPV/r alone for the treatment of COVID-19 patients (Deng et al., 2020).
Animals ; COVID-19/drug therapy* ; Drug Interactions ; Drug Therapy, Combination ; Female ; Indoles/pharmacokinetics* ; Lopinavir/pharmacokinetics* ; Male ; Rats ; Retrospective Studies ; Ritonavir/pharmacokinetics* ; SARS-CoV-2

Objective:To explore the epidemiological characteristics and the antibiotic resistance of Streptococcus pneumoniae isolates, and to provide the evidence for the rational use of antimicrobial agents to treat Streptococcus pneumoniae infection. Methods:The positive microbiological laboratory identification and antimicrobial susceptibility testing of Streptococcus pneumoniae from sputum of children with respiratory infections during January 2010 to December 2017 in Children′s Hospital of Soochow University were retrospectively analyzed. The positive rates of Streptococcus pneumoniae of different genders, ages, years and seasons were compared. The annual detection rates and trends of drug resistance of Streptococcus pneumoniae to penicillin, amoxicillin and cefotaxime were analyzed by Mann-Kendall trend test. The seasonal decomposition of time series was conducted to assess the association between Streptococcus pneumoniae detection rate and season. Enumeration data was compared using χ2 test. Results:Of the 88 480 sputum specimens, the total positive rate of Streptococcus pneumoniae was 10.3%(9 081/88 480). The detection rates of Streptococcus pneumoniae in children aged 0 to <0.5 years old, 0.5 to <2 years old, 2 to <3 years old, 3 to <5 years old, and 5 to <15 years old were 4.2%(1 407/33 224), 13.1%(3 191/24 390), 14.9%(2 417/16 252), 17.9%(1 474/8 246) and 9.3%(592/6 368), respectively. The difference was statistically significant ( χ2=2 421.6, P<0.01). The detection rates were 8.1%(1 321/16 306) from January to March, 10.9%(2 194/20 207) from April to June, 8.5%(2 141/25 058) from July to September, and 12.7%(3 425/26 909) from October to December. The discrepancy of positive rates in different seasons showed statistical significance ( χ2=311.5, P<0.01). During 2010 to 2017, significant decreases in antibiotic resistant rates of Streptococcus pneumoniae to penicillin, amoxicillin and cefotaxime were detected (tau=-0.93, -0.93 and -0.71, respectively, all P<0.05). Conclusions:The detection rate of Streptococcus pneumoniae in sputum of children with respiratory infections may present seasonal pattern and vary between different ages of children. The resistance to β-lactam antibiotics has declined.

Objective:To analyze the mean levels of skeletal muscle mass and strength in elderly patients with type 2 diabetes mellitus(T2DM), and to investigate the effects of chronic inflammatory factors and oxidative stress on them.Methods:A cross-sectional study was conducted on 120 patients with T2DM aged over 60 years and 126 elderly patients without diabetes(the control group). Skeletal muscle mass, strength and serum levels of chronic inflammatory factors interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and 8-hydroxy-2′-deoxyguanosine(8-OHdG)were determined, and their effects on skeletal muscle mass and strength in elderly patients with T2DM were analyzed.Results:Compared with the control group, grip strength decreased in elderly patients with T2DM(25.03±7.85)kg vs.(29.52±7.73)kg( P<0.01), and skeletal muscle mass decreased(21.36±5.46)kg vs.(22.01±5.22)kg with no significant difference( P>0.05). Serum levels of 8-OHdG were higher in elderly patients with T2DM than in the control group(3.08±0.26)ng/L vs.(2.59±0.16)ng/L( P<0.01). Correlation and regression analysis results showed that 8-OHdG was an influencing factor for muscle strength in elderly patients with T2DM( R2=0.457)and that height and weight could be influencing factors for skeletal muscle mass in elderly patients with T2DM( R2=0.822). Conclusions:Skeletal muscle mass and strength decline in elderly T2DM patients, probably as a result of increased levels of oxidative stress.These findings may serve as evidence for sarcopenia intervention in elderly T2DM patients.

Objective: Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province. Methods: A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data. Results: The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05). Conclusions The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.

Objective:To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon α-2b (rIFNα-2b) and the combination of lopinavir/ritonavir plus rIFNα-2b for patients with COVID-19 in Zhejiang province.Methods:A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNα-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data.Results:The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2±4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0±5.0) d] ( t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] ( H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively ( Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8±3.9), (13.5±5.1) and (11.2±4.3) d, respectively( Z=6.722, P<0.05). Conclusions:The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNα-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy; and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.

Objective To design a mathematical calculation model for better understanding and grasping the logical problem of replacement fluid and citric acid anticoagulant infusion in continuous veno-venous hemofiltration (CVVH). Methods ① Parameter definition: A, B, and T were respectively called the main part of pre-replacement fluid, 5% sodium bicarbonate solution, and 4% sodium citrate infused before filter. And a and b were respectively called the main part of post-replacement fluid, and 5% sodium bicarbonate solution infused after filter. ② Logic conversion:The liquid in back terminal (Z) was artificially divided into two parts. One (X) was the original residual plasma after filtration. The second (Y) was the part excluding the plasma, including the left part of pre-replacement fluid with sodium citrate, and the post-replacement fluid. ③The mathematical formulas of liquid volume and electrolyte concentration at X, Y and Z in unit time were listed according to the principle of CVVH and the screening coefficient of filter for different substances. ④The calculation formulas were entered into Excel form, and a mathematical calculation model was made, and a simulation calculation with examples was carried out. Results An Excel model was established by inserting the calculation formulas of volume, electrolyte, and total calcium at X, Y and Z. And it was found that the concentration of Na+, K+, Cl-, HCO3- at Y point remained unchanged only when A, B and (or) a, b was kept in same side and proportion even with the change of blood flow and other parameters without sodium citrate as anticoagulant. Once any of the parameters (such as blood flow, replacement fluid volume, etc.) were adjusted in other infusion methods (such as different ratios, different directions of the same year, etc.), the calculation results at Y would vary, and the electrolyte concentration at Z would change accordingly. A change of dilution model or parameter would result in the change of the electrolyte concentration at Y and Z with sodium citrate as anticoagulant. The concentration of total calcium scarcely changed no matter in what model and parameters. Conclusions All kinds of infusion ways could be included in the Excel model. The infusion results of all kinds of infusion matching could be intuitively evaluated. It is helpful for the medical staff to make a logical analysis and risk prediction in CVVH.