Objective:To evaluate the performance of an artificial intelligent (AI)-based automated digital cell morphology analyzer (hereinafter referred as AI morphology analyzer) in detecting peripheral white blood cells (WBCs).Methods:A multi-center study. 1. A total of 3010 venous blood samples were collected from 11 tertiary hospitals nationwide, and 14 types of WBCs were analyzed with the AI morphology analyzers. The pre-classification results were compared with the post-classification results reviewed by senior morphological experts in evaluate the accuracy, sensitivity, specificity, and agreement of the AI morphology analyzers on the WBC pre-classification. 2. 400 blood samples (no less than 50% of the samples with abnormal WBCs after pre-classification and manual review) were selected from 3 010 samples, and the morphologists conducted manual microscopic examinations to differentiate different types of WBCs. The correlation between the post-classification and the manual microscopic examination results was analyzed. 3. Blood samples of patients diagnosed with lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative neoplasms were selected from the 3 010 blood samples. The performance of the AI morphology analyzers in these five hematological malignancies was evaluated by comparing the pre-classification and post-classification results. Cohen′s kappa test was used to analyze the consistency of WBC pre-classification and expert audit results, and Passing-Bablock regression analysis was used for comparison test, and accuracy, sensitivity, specificity, and agreement were calculated according to the formula.Results:1. AI morphology analyzers can pre-classify 14 types of WBCs and nucleated red blood cells. Compared with the post-classification results reviewed by senior morphological experts, the pre-classification accuracy of total WBCs reached 97.97%, of which the pre-classification accuracies of normal WBCs and abnormal WBCs were more than 96% and 87%, respectively. 2. The post-classification results reviewed by senior morphological experts correlated well with the manual differential results for all types of WBCs and nucleated red blood cells (neutrophils, lymphocytes, monocytes, eosinophils, basophils, immature granulocytes, blast cells, nucleated erythrocytes and malignant cells r>0.90 respectively, reactive lymphocytes r=0.85). With reference, the positive smear of abnormal cell types defined by The International Consensus Group for Hematology, the AI morphology analyzer has the similar screening ability for abnormal WBC samples as the manual microscopic examination. 3. For the blood samples with malignant hematologic diseases, the AI morphology analyzers showed accuracies higher than 84% on blast cells pre-classification, and the sensitivities were higher than 94%. In acute myeloid leukemia, the sensitivity of abnormal promyelocytes pre-classification exceeded 95%. Conclusion:The AI morphology analyzer showed high pre-classification accuracies and sensitivities on all types of leukocytes in peripheral blood when comparing with the post-classification results reviewed by experts. The post-classification results also showed a good correlation with the manual differential results. The AI morphology analyzer provides an efficient adjunctive white blood cell detection method for screening malignant hematological diseases.

Objective: To explore the buffering effect of positive childhood experiences (PCEs) on mental health risks among adolescents before and after COVID-19 epidemic. Methods: In October 2019 (before the outbreak of COVID-19), 1 322 students from grades 4 to 9 were recruited from primary and secondary schools in two counties of Chizhou city, Anhui Province. A questionnaire survey was conducted to collect general demographic information, PCEs, depressive symptoms, anxiety symptoms, self harm behavior, suicidal ideation. Follow up survey was conducted after school re opening (May 2020). Mental health status before and after the COVID-19 epidemic was compared among students with different PCEs by multiple logistic regression analyses. Results: The detection rates of depressive symptoms, anxiety symptoms, self harm behavior and suicidal ideation (22.6%, 16.0%, 40.0%, 29.9%) of the respondents after school re opening were significantly higher compared that before the epidemic (16.5%, 13.5%, 31.1 %, 22.6%). There were no significant differences in the detection rates of depressive symptoms, anxiety symptoms,self harm behavior and suicidal ideation between high PCEs group before and after the epidemic ( Z =-0.05,0.27,0.84,1.84, P >0.05). Multivariate Logistic regression analysis showed that the risk of depressive symptoms and self harm behavior in the low PCEs group after school re opening was 1.39 times higher than that before the epidemic (95% CI= 1.05 -1.84, P <0.05). The risk of non suicidal self injury behavior in the low PCEs group after school re opening was 1.31 times higher than that before the epidemic (95% CI= 1.05 -1.62, P <0.05). There were no significant differences in mental health detection rates in high PCEs group before and after the epidemic ( P >0.05). Conclusion During the time of COVID-19 epidemic, PCEs is associated with lower rates of depressive symptoms, anxiety symptoms, self harm behavior and suicide ideation in adolescents. The findings suggest that more support and help should be given to adolescents from the perspectives of family, school and peers, so as to reduce the adverse effects of public health emergencies on adolescents mental health.

Objective: To investigate the epidemiological characteristics of premature eruption of permanent molars and its aasociation with body mass index (BMI), to provide a reference for childhood oral health promotion. Methods: A total of 861 children aged 9 to 12 years from two primary schools in Bengbu City were selected by cluster sampling method. Parental questionnaire was administered to collect socio demographic information. The eruption of second permanent molars were examined. Data was analyzed by multivariate Logistic regression model and margins command. Results: The detection rate of premature eruption of second permanent molars was 26.5%(228), 27.5% in boys and 24.9% in girls( χ 2=0.73, P =0.39). Early detection rate of second permanent molars (39.0%) was significantly higher in obese group than normal weight group (21.5%)( χ 2=21.85, P <0.01). Logistic regression analysis showed that obesity was positively correlated with the risk of premature eruption of second permanent molars( OR= 3.55 , 95%CI=2.14-5.87, P <0.01). Overweight was not associated with higher risk of premature eruption of second permanent molars( OR=1.64, 95%CI=0.95-2.81, P =0.07). Being female was associated with higher risk of premature eruption of second permanent molars compared to age matched peers( OR=2.19, 95%CI=1.42-3.39, P <0.01). Conclusion Childhood obesity is associated with higher risk for premature eruption of second permanent molars. Girls are more likely to have second permanent molar erupted in advance compared to age matched boys.

Objective:To establish an animal model of chronic systemic inflammation with long-term high expression of circulating IL-6 by introducing exogenous IL-6 gene transfer vector.Methods:Recombinant murine IL-6-encoding adeno-associated virus (AAV-IL-6) was constructed. Twenty-one 24-week-old male C57BL/6J mice were randomly divided into three groups with seven in each group: AAV-IL-6 group, vector control (AAV-ctrl) group and blank control group. At 0, 8 and 16 weeks of intervention, the mice in the three groups were injected with AAV-IL-6 (100 μl 0.5×10 10 vp/ml), unloaded AAV (100 μl 0.5×10 10 vp/ml) and the same volume of saline in the tail vein, respectively. IL-6 levels in mouse serum were measured by ELISA. The general condition of mice was observed and blood routine tests were performed. Changes in blood biochemical parameters and C-reactive protein (CRP) levels were detected. At the end of 24-week intervention, the mice were sacrificed and the myocardium, liver, spleen, quadriceps femoris, knee joint and middle femur were taken for HE staining. Results:At 4, 8, 16 and 24 weeks after intervention, serum IL-6 levels were (75.41-169.28) pg/ml in the AAV-IL-6 group, while in the two control groups, the levels were below the lower limit of detection (7.8 pg/ml). At 24 weeks after intervention, the body weight of mice in the AAV-IL-6 group was significantly lower than that of mice in the two control groups; the neutrophil counts and CRP level in the AAV-IL-6 group were higher than those in the two control groups, while the levels of albumin, creatinine, triglyceride and cholesterol were lower than those in the two control groups. There were no differences in the aforementioned parameters between the two control groups. Compared with the blank control group, both AAV-IL-6 and AAV-ctrl groups showed increased lymphocyte counts. All mice had normal liver and kidney functions at the end of intervention. Histopathological findings indicated that the mice in the AAV-IL-6 group had focal infiltration of lymphocytes in the central venous area of the liver and around the myocardial and the skeletal muscle fibers, diffuse infiltration of multinucleated giant cells in the spleen, atrophic skeletal muscle, disorganized growth plate, reduced chondrocyte hypertrophic zone, thinner bone cortex and trabecular, and reduced osteoid. There were no histopathological changes in mice of the two control groups.Conclusions:Repeated tail vein injection of AAV-IL-6 could achieve long-term high expression of circulating IL-6 in mice, which manifested the phenotype of chronic systemic inflammation in preliminary detection and provided a safe, effective and simply accessible animal model for related studies.

OBJECTIVE：To prepare Imperatorin ultradeformable liposomes gel （IMP-UDLs-Gel），and to evaluate its quality. METHODS：Based on single factor test ，using 12 h accumulative penetration amount （Q12h）as evaluation index ，the proportion of carbomer 940，glycerol and propyl glycol in formulation of IMP-UDLs-Gel were investigated by orthogonal test. The optimal formulation was screened. The quality of IMP-UDLs-Gel prepared with the optimal formulation was evaluated. RESULTS ：The optimal formulation of IMP-UDLs-Gel included carbomer 940 proportion of 1％，glycerol proportion of 15％ and propyl glycol proportion of 10％. Q12 h of IMP-UDLs-Gel was （11.543±0.241）μg/cm2；the appearance was milky white and translucent ；the particle size was （93.13±1.68）nm，PDI was 0.268±0.012，Zeta potential was （－24.96±1.99）mV；pH was 7.32±0.03； viscosity was （45.37±1.27）g·s；steady flow was （0.727±0.002）μg·h/cm2，lag time was （4.358±0.175）h，apparent permeability coefficient was 1.392×10－3 cm/h，and it has good physical and optical stability. CONCLUSIONS ：The preparation method is stable and feasible ，and the prepared IMP-UDLs-Gel has good adhesion ，stability and transdermal property.

Objective To understand the types and distribution of Arboviruses in Hainan province.Methods Blood-sucking insects were collected in Hainan province from 2017 to 2018.After laboratory treatment,BHK-21 cells and C6/36 cells were inoculated with grinding supernatant of all blood-sucking insects to isolate all of involving virus.Arbovirus genes in blood-sucking insects were detected in parallel by RT-PCR method.Results A total of 15 062 mosquitoes were classified into four genera (Culex,Armigeres,Aedes,Anopheles) and 11 360 midges were collected.Culex tritaeniorhynchus was in the majority and accounted for 92.88％ (13 990/15 062) of all the mosquitoes collected.Four strains of virus isolates were notified by tissue culture method.Three strains of viruses belonged to Japanese encephalitis virus (JEV),with the other one as Getah virus (GETV).Five pools of JEV gene amplification were positive,from Culex tritaeniorhynchus.Results from the phylogenetic analysis showed that they belonged to genotype JEV-Ⅰ.The minimum infection rate of JEV was 0.57‰ (8/13 990).A total of 5 pools of Akabane virus (AKV) gene amplification were positive.The minimum infection rate of AKV was 0.44‰ (5/11 360).Based on the S gene and M gene sequences of the virus,data from the phylogenetic analysis showed that the five AKV strains carried by midges in Hainan province were in a separate evolutionary branch and with formed unique geographical distribution.Conclusions JEV and GETV had been isolated again from the mosquito specimens in this survey,since the 1980s.AKV was detected from the midge specimens in Hainan province.These results showed the needs of strengthening the programs on detection and monitor of JEV,GETV and AKV that were related to animal and human diseases in order to reduce the risks of related diseases in this area.

Objective To investigate the correlation of ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,AluⅠ digestion polymorphism with coronary atherosclerotic heart disease(CHD) risk factors in Yancheng area .Methods A total of 124 cases of CHD and 163 persons undergoing physical examination served as the CHD group and CON group .The enzyme method was adopted to detect TG and TC .The direction method was adopted to detect HDL and LDL .ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,AluⅠ digestion polymorphisms were detected by adopting RFLP-PCR .Results The ratios of smoking history ,family history ,complicating hypertension and diabetes ,and the level of body mass index ,TC ,TG and LDLC in the CHD group were significantly higher than those in the control group ,the difference was statistically significant (P＜0 .05) .The various indicators had no statistically difference between male and female(P＞0 .05) .The frequency distribution and geographic distribution of ERα gene PvuⅡ ,XbaⅠ and ERβ gene RsaⅠ ,Alu Ⅰ digestion polymorphisms had no difference between the two groups ,all conformed to Hardy-Weinberg genetic equilibrium and had the group representativeness .pp ,xx ,RR and AA genotypes in the CHD group were maximal ,while PP , XX ,rr and aa genotypes were minimal ;Pp ,xx ,RR and AA genotypes in the CON group were maximal ,while PP ,XX ,rr and aa genotypes were minimal .The distribution frequency of p and x genes in the CHD group was significantly higher than that in the control group ,the difference was statistically significant(P＜0 .05) .Conclusion The estrogen gene polymorphism might be a target spot for effectively treating CHD ,and p and x gene distribution frequency may be related with CHD risk factors .

Background Immediately ocular rinsing is a key step for the prevention of eye tissue damage after acute chemical bums.A good ophthalmic rinsing solution can neutralize chemical substance and improve the prognosis of patients.Objective This study attempted to evaluate the therapeutic effect of self-made citric aciddisodium hydrogen phosphate buffer and citric acid-disodium hydrogen phosphate-potassium chloride buffer on corneal chemical burns in mice.Methods Citric acid-disodium hydrogen phosphate buffer solution (solution 1) and citric acid-disodium hydrogen phosphate-potassium chloride buffer solution (solution2) with the pH 7.4 were prepared.One hundred and twenty clean male C57 mice aged 6-8 weeks were randomized to two groups,and filter papers containing 1 mol/L H2SO4 or 0.15 mol/L NaOH were attached to the central corneas of the right eyes to create the acid or alkali burning models.Then the eyes were immediately rinsed by 40 ml solution 1,tap water or solution 2 according to the grouping and the model eyes without rinsing served as the control group.The corneal opacity was examined by slit lamp microscope and scored in 3,7 and 14 days after modeling.The percentages of corneal fluorescein staining,corneal neovascularization and corneal ulcer were analyzed.The study protocol was approved by Experimental Animal Ethic Commission of Second Military Medical University.Results In the corneal acid burning models,the number of eye with corneal opacity scored 1 in the solution 1 group,tap water group and solution 2 group was significantly more than that in the non-rinsing group in 3,7 and 14 days after modeling (all at P＜0.01);In 3 days after modeling,the numbers of eye scored 1 were more in the solution 1 group than those in the tap water group and solution 2 group (x2 =11.000,P =0.001;x2 =4.000,P =0.046).There were no differences in the eye number of different corneal opacity scores in 14 days after acid burning (all at P＞0.05).In 3,7 and 14 days after corneal alkali burning,the number of eyes with corneal opacity scored 1-2 was significantly increased in the solution 1 group,tap water group and solution 2 group compared with non-rinsing group (all at P＜0.01).The percentage of corneal ulcer in the solution 1 group,tap water group and solution 2 group was 7％,27％ and 13％,respectively,which was significantly lower than 73％ in the non-rinsing group (P =0.000,0.027,0.003),and no significant differences were seen in various time points after corneal alkali burning (all at P＞O.05).Corneal neovascularization occurred in 50％ mice in non-rinsing group in 14 days after acid burning.However,no neovascularization was seen in the mice of the solution 1 group,tap water group and solution 2 group in both acid and alkali burning mice.Conclusions Citric aciddisodium hydrogen phosphate buffer (pH 7.4) appears to be an effective emergency rinsing solution for corneal chemical burns and the rinsing solution with or without potassium chloride is not obviously affected to the prognosis of corneal chemical burns in the mice.

Objective:To evaluate the clinical applicability of CT angiography (CTA) for locating the perforator vessels of anterolateral thigh flap(ALTF) and the effects of individualized ALTF designed by CTA for the reconstruction of the soft tissue defects of tongue after the resection of tongue carcinoma.Methods:21 patients with tongue squamous cell carcinoma underwent CTA for locating the perforator vessels of ALTFs and for the design of individualized ALTFs before operation.The patients underwent soft tissue defect reconstruction with individualized ALTFs after tumor removal.Results:All the operations came off as preoperative designed,the intraoperative findings of the blood vessel alignment were consistent with the preoperative CTA results.The size of flaps was 6.0 cm × 5.0 cm-11.0 cm× 8.5 cm.20 ALTFs survived,2 ALTFs appeared vascular crisis,1 remained survival and the other was necrotic after surgical exploration.During 6 ～ 60 momhs of follow-up,the survival condition of flaps and the wound healing condition were both satisfactory.1 patient died because of distant metastasis 18 months after operation.20 patients reminded free of carcinoma and satisfied with the reconstructive effects of chewing,swallowing and linguistic function.Conclusion:CTA can accurately locate perforator vessels for the design of individualized AFLT.AFLT designed by CTA is an ideal choice for the reconstruction of postoperative soft tissue defects after resection tongue carcinoma.

Objective To investigate the relationship between adiponectin combined with the ultrasound blood flow index of the umbilical artery and perinatal outcome in women with severe preeclampsia. Methods Placental tissues were obtained from normal term pregnancies (control group, n=50) and severe preeclampsia patients (PE group, n=50) in Third Affiliated Hospital of Zhengzhou University from February 2014 to October 2014. The expression of adiponectin was examined using immunohistochemical methods and real-time polymerase chain reaction. The umbilical artery was measured by color Doppler, and the umbilical artery systolic/diastolic ratio (UA-S/D), umbilical artery resistance index (UA-RI) and umbilical artery pulsatility index (UA-PI) were determined. The relationship between the expression of adiponectin in placental tissues, UA-S/D and perinatal outcome were analyzed. The data were analyzed using two dependent-sample t test, the log-rank test and Spearman correlation analysis. Results Compared with the control group, infants in the PE group had lower birth weight and placental weight, shorter height, and greater umbilical artery indices including UA-S/D, UA-RI and UA-PI (all P<0.05). The expression of adiponectin and its mRNA in placentae of the PE group was significantly higher than that of the control group (adiponectin: 0.326±0.011 vs. 0.116±0.011, t=99.144, P=0.000;mRNA:4.18±1.80 vs. 1.00±0.51, t=11.985, P=0.000). UA-S/D had a negative correlation with birth weight, onset gestational age and gestational age at birth (r= - 0.897, - 0.469 and - 0.524, all P<0.01). The expression of adiponectin mRNA had a negative correlation with birth weight, onset gestational age, and gestational age at birth (r=-0.580,-0.407 and-0.449, all P<0.01). The expression level of adiponectin had positive correlations with body mass index of the mothers and the UA-S/D (r=0.261 and 0.788, both P<0.01). Conclusions The expression of adiponectin in placental tissues and blood flow index of the umbilical artery both increase in severe preeclampsia, and are associated with poor perinatal outcome.