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 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.

OBJECTIVES: Acute kidney injury (AKI) can be caused by ischemia/reperfusion (I/R), nephrotoxin, and sepsis, with poor prognosis and high mortality. Leptin is a protein molecule that regulates the body's energy metabolism and reproductive activities via binding to its specific receptor. Leptin can inhibit cardiomyocyte apoptosis caused by I/R, but its effect on I/R kidney injury and the underlying mechanisms are still unclear. This study aims to investigate the effect and mechanisms of leptin on renal function, renal histopathology, apoptosis, and autophagy during acute I/R kidney injury. METHODS: Healthy adult male mice were randomly divided into 4 groups: a sham+wild-type mice (ob/+) group, a sham+leptin gene-deficient mice (ob/ob) group, an I/R+ob/+ group, and an I/R+ob/ob group (n=8 per group). For sham operation, a longitudinal incision was made on the back of the mice to expose and separate the bilateral kidneys and renal arteries, and no subsequent treatment was performed. I/R treatment was ischemia for 30 min and reperfusion for 48 h. The levels of BUN and SCr were detected to evaluate renal function; HE staining was used to observe the pathological changes of renal tissue; TUNEL staining was used to observe cell apoptosis, and apoptosis-positive cells were counted; Western blotting was used to detect levels of apoptosis-related proteins (caspase 3, caspase 9), autophagy-related proteins [mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), LC3 I, LC3 II], mTOR-dependent signaling pathway proteins [phosphate and tension homology (PTEN), adenosine monophosphate-activated protein kinase (AMPK), protein kinase B (AKT), extracellular regulated protein kinase (ERK), phosphorylated PTEN (p-PTEN), phosphorylated AMPK (p-AMPK), phosphorylated AKT (p-AKT), phosphorylated ERK (p-ERK)]. RESULTS: There was no significant difference in the levels of BUN and SCr between the sham+ob/+ group and the sham+ob/ob group (both P>0.05). The levels of BUN and SCr in the I/R+ob/+ group were significantly higher than those in the sham+ob/+ group (both P<0.05). Compared with the mice in the sham+ob/ob group or the I/R+ob/+ group, the levels of BUN and SCr in the I/R+ob/ob group were significantly increased (all P<0.05). There was no obvious damage to the renal tubules in the sham+ob/+ group and the sham+ob/ob group. Compared with sham+ob/+ group and sham+ob/ob group, both the I/R+ob/+ group and the I/R+ob/ob group had cell damage such as brush border shedding, vacuolar degeneration, and cast formation. Compared with the I/R+ob/+ group, the renal tubules of the mice in the I/R+ob/ob group were more severely damaged. The pathological score of renal tubular injury showed that the renal tubular injury was the most serious in the I/R+ob/ob group (P<0.05). Compared with the sham+ob/+ group, the protein levels of caspase 3, caspase 9, PTEN, and LC3 II were significantly up-regulated, the ratio of LC3 II to LC3 I was significantly increased, and the protein levels of p-mTOR, p-PTEN, p-AMPK, p-AKT, and p-ERK were significantly down-regulated in the I/R+ob/+ group (all P<0.05). Compared with the sham+ob/ob group, the protein levels of caspase 3, caspase 9, PTEN, and LC3 II were significantly up-regulated, and the ratio of LC3 II to LC3 I was significantly increased, while the protein levels of p-mTOR, p-PTEN, p-AMPK, p-AKT, and p-ERK were significantly down-regulated in the I/R+ob/ob group (all P<0.05). Compared with the I/R+ob/+ group, the levels of p-mTOR, p-PTEN, p-AMPK, p-AKT were more significantly down-regulated, while the levels of caspase 3, caspase 9, PTEN, and LC3 II were more significantly up-regulated, and the ratio of LC3 II to LC3 I was more significantly increase in the I/R+ob/ob group (all P<0.05). CONCLUSIONS Renal function and tubular damage, and elevated levels of apoptosis and autophagy are observed in mice kidneys after acute I/R. Leptin might relieve I/R induced AKI by inhibiting apoptosis and autophagy that through a complex network of interactions between mTOR-dependent signaling pathways.
AMP-Activated Protein Kinases/metabolism* ; Acute Kidney Injury/pathology* ; Animals ; Apoptosis ; Apoptosis Regulatory Proteins/pharmacology* ; Autophagy ; Caspase 3/metabolism* ; Caspase 9/metabolism* ; Female ; Humans ; Ischemia ; Kidney/pathology* ; Leptin/pharmacology* ; Male ; Mammals/metabolism* ; Mice ; Proto-Oncogene Proteins c-akt/metabolism* ; Reperfusion/adverse effects* ; Reperfusion Injury/metabolism* ; TOR Serine-Threonine Kinases/metabolism*

Objective:To predict the financial impact of artificial intelligence (AI)-assisted community-based screening for primary angle-closure glaucoma (PACG) among the elderly on health expenses and medical insurance expenditures in remote areas of China.Methods:A total of 19 395 people aged more than 65 years old from Changjiang Li autonomous county, Hainan province were enrolled.A health economic model for glaucoma screening in Wenzhou, Zhejiang province was adopted.PACG was classified as suspected primary angle closure, primary angle closure, PACG, monocular blindness and binocular blindness according to its severity.The model was adjusted in combination with the epidemiology, metastasis probability and treatment compliance, screening and referral compliance, sensitivity and specificity of AI screening from glaucoma related literature and using data on local epidemiology and health costs of glaucoma from the Changjiang Hospital of Integrated Traditional Chinese and Western Medicine from 2016 to 2019.The health outcome, total health expenses and medical insurance expenditures of AI-assisted community-based screening and no screening for PACG in 15 years were compared and analyzed.Results:It is found that the AI-assisted community-based screening would prevent disease progression and reduce the number of PACG patients by 43% (134 cases) and blind patients by 50% (9 cases) in 15 years in comparison with no screening.Under the AI-assisted community-based screening system, the early diagnosis and intervention of positive PACG patients will cause the dramatic increase of both total health expenses and medical insurance expenditures compared with no screening in the short term.It was predicted that the increasing trend of medical insurance expenditure would turn to a downward trend and show cost saving advantage at the third year, whereas the declining trend of total health expense would not occur until 10 years later.Without screening, a large number of PACG patients would progress to advanced stages, so the medical costs would continue to rise and exceed the medical costs of the screening system at the 14th year.Conclusions:The AI-assisted community-based screening for PACG is cost-effective and can prevent disease progression, save health expenses and medical insurance expenditures in the long run in remote areas of China.

Objective:To evaluate the clinical significance of our self-designed three-column scoring system for postoperative X-ray stability of intertrochanteric fracture after intramedullary nailing.Methods:A retrospective study was conducted of the 378 patients with intertrochanteric fracture who had been treated between January 2015 and June 2019 at Department of Orthopaedics, Linyi People's Hospital by internal fixation with proximal femoral nail antirotation (PFNA). They were 161 males and 217 females, aged from 60 to 97 years (average, 72.5 years). By the AO classification, there were 109 cases of type 31-A1, 188 cases of type 31-A2 and 81 cases of type 31-A3. Anteroposterior and lateral X-ray examinations of the hip were performed immediately after internal fixation to evaluate the fracture stability using our self-designed three-column scoring system by which the medial column is given 4 points, middle column 2 points and lateral column 2 points. A single column scoring full points is rated as stable, 3-column stability as excellent, 2-column stability as good, one-column stability as fair, and 3-column unstability as poor. Rehabilitation programs were carried out according to the results of stability evaluation: full weight bearing at an early stage was indicated for excellent patients, partial weight bearing at an early stage for good patients and weight bearing at an early stage contraindicated for fair or poor patients. Fracture union time and failure of internal fixation were recorded. The relationship between internal fixation failure and our three-column scoring system was calculated.Results:The 378 patients were followed up for 6 to 24 months (mean, 10.4 months). Of them, 365 obtained fracture union after an average time of 4.3 months (from 3 to 7 months). Internal fixation failure occurred in 13 patients, giving a failure rate of 3.4%(13/378).Of the 129 excellent patients by our three-column scoring system for post-operative X-ray stability of intertrochanteric fracture, none failed in internal fixation; of the 193 good patients, 4 failed; of the 56 fair patients, 9 failed. Internal fixation failure was closely related to our three-column scoring system for postoperative X-ray stability of intertrochanteric fracture ( r=-0.986, P=0.006), as well as to the column stability ( r=-1.000, P=0.033). Conclusion:Our self-designed three-column scoring system for postoperative X-ray stability of intertrochanteric fracture after intramedullary nailing can accurately reflect the fracture stability so that it can be used to guide rehabilitation programs for the patients and judge their prognosis.

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 analyzed demographic and clinical characteristics in hospitalized patients with osteoporotic vertebral compression fractures(OVCFs)and to provide certain evidence for prevention and treatment of the disease. Methods: Retrospectively analyzedthe clinical data of 2 028 patients with OVCF from January 2009 to December 2018 in the Spine Department of Qilu Hospital of Shandong University. The trend of composition ratio and average age of OVCF patients in the past ten years were observed, the differences of the demographic characteristics and clinical characteristics between different genders as well as the correlation between the number of fractured vertebral bodies of OVCF and various influencing factors were analyzed, and the characteristics of OVCF inpatients were summarized. Results: Of 2 028 patients, 374 were males and 1654 were females.The average age was 70.50±9.73 years old, and the male is 73.29±10.23 and the female is 69.87±9.46. A significant increase in the constituent ratio of OVCF between 2009 and 2018 (6.85% in 2009 vs. 13.24% in 2013) and the mean age of OVCF patients increased from 67.54±9.92 years in 2009 to 70.96±9.63 years in 2018were identified.There were significant differences in age distribution of OVCFs in different gender. When the age was less than 70 years old,the fracture ratio of female was higher than that of males, especially between 60 and 70 years old (38.63%∶17.91%); However, when the age was more than 70 years, the fracture ratio of male was higher than that of female especially after 80 years old(29.15%:12.70%). There were significant differences in occupation distribution of OVCFs indifferent gender. Peasants (47.94%), professionals (18.32%) and workers (14.63%) were the top three in female while peasants (35.56%), workers (26.47%) and professionals(20.05%) in male. Falling was the main injury cause of OVCF and there were significant differences in the distribution of causes of OVCF injuries between different genders. The proportion of traffic accident injuries infemaleswas higher than that in maleswhile the proportion of sprains inmaleswas higher than that in females.Moreover,there was a statistically significant difference in the distribution of fracture vertebral bodies between patients of different ages, with or without fracture history and smoking history and different ages of menopause.Among them, the number of fractured vertebral bodies was positively correlated with age(r=0.79, P<0.05).The composition ratio of single vertebral fracture of patients without fracture history, menopausal ageunder 50 and withoutsmoking history was higher than that with fracture history, menopausal age over 50 and with smoking history. On the contrary,the composition ratio of two or more vertebral fractures in patients without fracture history and menopausal agebelow 50 was lower than thatwith fracture history and menopausal ageover 50. Particularly, the ratio of three or more vertebral fractures with smoking history was higher than thatwithout smoking history.The number of patients who adopted bone mineral density(BMD) test and bone turnover maker test was 118 and 106 before 2014 while the number was 568 and 837 after 2014 and the differences were statistically significant. Conclusion The proportion of OVCFs in spine diseases and average age of patients in the period of 2009-2018 was increasing gradually. Distribution of the age,occupation and cause of injury were related to the gender, but there was no correlation between distribution of season, fracture segment and comorbidities and gender. The age, previous fracture history, female menopausal age and smoking history were correlated with the number of fractured vertebral bodies while drinking history was not related.

Objective To explore the risk factors for childhood death from pneumococcal meningitis.Methods The data of 32 hospitalized children were retrospectively analyzed,who were diagnosed as pneumococcal meningitis and enrolled in the Affiliated Children′s Hospital of Soochow University from November 201 0 to December 201 5.The subjects were divided into the death group and survival group according to their prognosis.The clinical characteristics and laboratory data were compared between 2 groups.Results Between the death group and survival group,there were significant statistically differences in shock within 24 hours after admission(63.6% vs 1 4.3%,P =0.01 3),as well as endotracheal tube intubation(1 00.0% vs 23.8%,P <0.001 ),the levels of cerebrospinal fluid(CSF)IgG[(491 .27 ± 203.53)mg/L vs (267.24 ±1 88.07)mg/L,P =0.006],IgM[(1 1 5.72 ±79.1 9)mg/L vs (32.80 ±28.52)mg/L, P =0.006],IgA[59.52(1 5.51 ,75.69)mg/L vs 1 8.77(9.33,27.54)mg/L,P =0.023],CSF leukocyte[330.00 (1 50.00,380.00)×1 06 /L vs 870.00 (403.00,6 1 60.00)×1 06 /L,P =0.009 ],CSF protein [(4 047.00 ± 1 942.1 6)mg/L vs (2 470.62 ±1 259.94)mg/L,P =0.009],CSF adenosine deaminase (ADA)[35.20(1 8.90, 87.20)U /L vs 8.80(3.05,23.78)U /L,P =0.001 ],serum sodium[(1 30.21 ±2.85)mmol/L vs (1 32.83 ±3.69) mmol/L,P =0.049],serum lactic acid (LA)[4.40 (2.60,5.70)mmol/L vs 2.40 (1 .75,4.50)mmol/L,P =0.01 3],serum C -reactive protein (CRP)[(95.87 ±65.40)mg/L vs (1 65.61 ±83.05)mg/L,P =0.022],serum lactate dehydrogenase (LDH)[81 3.40(465.20,2 31 0.70)U /L vs 359.20(257.85,405.90)U /L,P =0.001 ], platelet[(1 63.82 ±1 64.86)×1 09 /L vs (295.71 ±1 30.29)×1 09 /L,P =0.01 9]and positive rate of blood culture (90.9% vs 47.6%,P =0.023)between the death group and survival group.Conclusions The risk factors associated with mortality in pediatric SPM include shock within 24 hours after admission,endotracheal intubation,hyponatremia, thrombocytopenia,as well as high serum LA level,high serum LDH level,lower serum CRP level or cultures of blood and CSF double positive.

Objective To study the accuracy of the anteversion of femoral prosthesis in hemiarthro-plasty for treating femoral intertrochanteric fracture in the elderly.Methods Twenty femoral neck antever-sions of specimens were measured, which were control group(A).The femoral specimens were made into the model of femoral intertrochanteric fracture, which were installed femoral prosthesis according to minor trochanter(B),identifying the max diameter of cavum medulla(C),ocular estimating femoral condylar line ( D) ,combination of identifying the max diameter of cavum medulla and ocular estimating femoral condylar line( E) respectively.All the anteversion of femoral prosthesis were measured.Results The anteversion were:A group(8.98 ±4.31)°,B group (9.12 ±4.87)°,C group (9.57 ±6.49)°,D group (13.18 ± 5.83)°,E group (10.37 ±4.56)°respectively.The difference among the five groups was statistically sig-nificant.There was difference between D and A, there were no differences between B and A, C and A, E and A respectively.Conclusions In hemiarthroplasty for treating femoral intertrochanteric fracture, the an-teversion of femoral prosthesis is according to minor trochanter possibly; if the minor trochanter can not be got reduction, it is a good option according to combination of identifying the max diameter of cavum medulla and ocular estimating femoral condylar line to make sure the accuracy of the anteversion of femoral prosthe-sis.