Objective:To analyze and compare the differentially expressed genes (DEGs) of Yes-associated protein (YAP)-positive and negative hepatocytes and further understand the preliminary functional characteristics of YAP-positive hepatocytes in an early mouse model of nonalcoholic steatohepatitis (NASH) with transcriptome sequence (RNA-Seq).Methods:C57BL/6 mice were fed with methionine-choline deficiency (MCD) diet for 2 weeks to establish an early NASH model, and the control group was fed with normal diet. Liver tissue was stained with hematoxylin-eosin (HE) and Sirius red, and the pathological score was recorded. The expression of YAP and P-YAP were determined by immunohistochemistry (IHC) in liver tissues. Primary hepatocytes with viability greater than 90% were isolated and purified by collagenase perfusion combined with Percoll density gradient centrifugation. YAP-positive and negative hepatocytes were assessed by YAP antibody, flow cytometry and RNA-Seq analyses. Sequencing results were screened by GO, KEGG and interaction network analysis methods. RT-PCR was used to verify the expression levels of YAP and some DEGs in liver tissue model group. Two samples mean was compared by independent samples t-test. Results:Compared with the control group, the HE-stained liver tissue of MCD-induced mice at 2 weeks showed steatosis (pathological score 1.07±0.21), accompanied by lobular inflammation (pathological score 1.13±0.32) and ballooned hepatocyte (pathological score 0.80) ±0.20). Sirius red staining showed non-significant liver fibrosis (pathological score 0.40±0.40). IHC showed partial YAP-positive hepatocytes expression in an early stage of NASH. RNA-Seq analysis showed that clean reads of YAP-positive and negative hepatocytes were 49 310 604 and 5 4820 036, respectively. Compared with YAP-negative hepatocytes, YAP-positive hepatocytes had differential expression of 5 565 genes, including 1 662 up-regulated genes and 3 903 down-regulated genes. GO analysis of up-regulated genes showed that the metabolic processes related to mitochondrial functions, such as purine nucleoside triphosphate and nucleoside triphosphate were significantly enriched in biological processes (BP), while down-regulated gene analysis showed that olfactory-related receptor were significantly enriched in BP. KEGG analysis showed that DEGs were enriched in 292 pathways, and oxidative phosphorylation (OXPHOS) pathway was significantly enriched in signaling pathway. RT-PCR validated that inflammatory factors (interleukin-1β, interleukin-6), YAP and its target genes (Cyr61, Ankrd1), and Cox5b and Sdhc genes were significantly up-regulated in the OXPHOS pathway, which was consistent with the sequencing results. In addition, eight key genes with interaction network analysis were predicted.Conclusion:Changes in hepatocyte metabolic levels may be associated with increased YAP activity in an early stage of NASH.

Objective:To investigate the effect of the adaptive computerized cognitive training(ACCT) on patients with mild cognitive impairment(MCI).Methods:A total of 114 patients with mild cognitive impairment (53 cases in the treatment group and 61 cases in the observation group) were selected.In the treatment group, routine treatment combined with ACCT were given for 24 weeks, then routine treatment only for 24 weeks, 48 weeks altogether. In the observation group, routine treatment was given for 48 weeks.At week 0, 24, 48, both groups were assessed by scales including: mini-mental state examination (MMSE), Montreal cognitive assessment(MoCA), numerical memory span test, activities of daily living, Hamilton depression scale(HAMD)and Hamilton anxiety scale(HAMA). SAS 9.4 was used for statistical analysis.The data were analyzed by χ 2 test, rank sum test and multi-level model analysis. Results:Interactions between group and time on MMSE( treatment group: 0 week 22.0 (21.0, 23.0), 24 weeks 24.0 (24.0, 25.0), 48 weeks 25.0 (24.0, 26.0); observation group: 0 week 23.0 (21.0, 24.0), 24 weeks 23.0 (21.0, 24.0), 48 weeks 23.0 (21.0, 24.0)), MoCA( treatment group: 0 week 18.0 (17.0, 20.0), 24 weeks 22.0 (20.0, 23.0), 48 weeks 22.0 (20.0, 24.0); observation group: 0 week 19.0 (17.0, 20.0), 24 weeks 19.0 (18.0, 20.0), 48 weeks 19.0 (18.0, 20.0)), IADL( treatment group: 0 week 11.0 (10.0, 13.0), 24 weeks 12.0 (10.0, 12.0), 48 weeks 12.0 (10.0, 12.0); observation group: 0 week 12.0 (11.0, 13.0), 24 weeks 11.0 (10.0, 12.0), 48 weeks 11.0 (10.0, 12.0)), DST-forwards and HAMD scores were significant(all P<0.05), and DST-backwards had significant group main effect ( P<0.05). Further simple effect analysis showed that the influence of group and time on MMSE, MoCA and DST-forwards were statistically significant (all P<0.05), and the influence of time on IADL and HAMD were statistically significant (both P<0.05). Further comparison of the difference between the two groups at each time point: D-value of MMSE, MoCA, DST-forwards, and DST-backwards score in the treatment group were higher than those in the observation group, while D-value of HAMD score was lower than that in the observation group, and the differences were statistically significant (all P<0.05). Conclusion:ACCT may have long term effect on improving the cognitive function and depression of MCI patients.

Objective:To investigate the clinical efficacy of finasteride combined with 5% minoxidil in the treatment of male androgenetic alopecia.Methods:A total of 102 males with androgenetic alopecia diagnosed and treated in Shaoxing Hospital of China Medical University during 2018-2019 were selected in the study and divided into two groups by random number table method.The control group (50 cases) was only given finasteride, and the treatment group(52 cases) was given finasteride combined with minoxidil.All the patients were treated for 6 months.The therapeutic effects and adverse reactions during the treatment were compared between the two groups.Results:The effective rate of the treatment group was 86.5%(45/52), which was significantly higher than 66.0%(33/50) of the control group(χ 2=4.93, P<0.05). The incidence of adverse reactions in the treatment group was 3.8%(2/52), which was significantly lower than 18.0%(9/50) in the control group(χ 2=5.22, P<0.05). Conclusion:Finasteride combined with 5% minoxidil is safe and effective in the treatment of male androgenetic alopecia, which is worthy of clinical promotion.

Objective:To observe the clinical efficacy of Danzhi Sangqin decoction in the treatment of premenstrual acne with liver and lung stagnation.Methods:Sixty patients with liver and lung stagnation premenstrual acne diagnosed and treated in the Department of Dermatology of Shaoxing Central Hospital from July 2016 to November 2018 were selected in the research.They were divided into the control group( n=28) and treatment group( n=32) by random number table method.The treatment group was treated with self-made Danzhi Sangqin decoction, and the control group was orally treated with isotretinoin capsules and vitamin B6.The two groups were treated for 1 week.The clinical efficacy and the incidence of adverse reactions were compared between the two groups. Results:In the treatment group, 18 cases were markedly effective, 11 cases were effective, and 3 cases were ineffective, and the total effective rate was 90.6%(29/32). In the control group, 16 cases were significantly effective, 10 cases were effective, and 2 cases were ineffective, and the total effective rate was 92.9%(26/28). There was no statistically significant difference in the total effective rate between the two groups( P>0.05). The incidence of adverse reactions in the treatment group was 3.1%(1/32), which was significantly lower than that in the control group [21.4%(6/28)], the difference was statistically significant(χ 2=6.281, P<0.05). Conclusion:The clinical efficacy of Danzhi Sangqin decoction in the treatment of premenstrual acne with liver and lung stagnation is similar to that of isotretinoin capsules, but the incidence of adverse reactions is low.

Objective: To apply the semi-quantitative risk assessment model in the Technical Guidelines for Occupational Health Risk Assessment of Chemical Hazardous Factors in the Workplace to assess the occupational health risk of electroplating enterprises and explore its applicable conditions and characteristics. Methods: Three electroplating enterprises were selected as the research objects. Occupational hygiene survey and hazard factor detection were conducted on the spot. Three semi-quantitative risk assessment methods were used to assess the risk, and the evaluation results were compared. Results: The consistency between the contact index method and the comprehensive index method was strong. The weighted Kappa value was 0.946 and the P value was less than 0.001, but the consistency between the contact ratio method, the contact index method and the comprehensive index method was poor. The weighted Kappa value was 0.345 and 0.391, and the P value was 0.009 and 0.004, respectively. When the contact concentration is less than 50% OELs, the evaluation results of the contact ratio method are lower than those of the exponential method and the comprehensive exponential method. The consistency of the three methods was the highest when the contact concentration (>50%) and (OELs, the results of the contact ratio method are higher than those of the index method and the comprehensive index method. Conclusion Contact ratio method is suitable for occupational health risk assessment under the condition of incomplete occupational health information and for enterprise managers to identify key control points of health risk through self-assessment; Contact index method is suitable for hazard risk assessment of occupational hazards without sampling test conditions or OELs, and pre-assessment of occupational hazards of construction projects without access to analogical test data. The index method is suitable for occupational health risk assessment with available testing data and complete occupational health information.

Objective To explore the value of controlled decompression under intracranial pressure monitoring in craniotomy of patients with severe cerebral hemorrhage.Methods One hundred and six patients with severe cerebral hemorrhage,admitted to our hospital from January 2015 to July 2018,were prospectively enrolled.These patients were divided into control group (n=5 l) and treatment group (n=55) according to their families' wishes.The patients in the control group were treated with traditional craniotomy and hematoma removal;the patients in the treatment group were treated with controlled decompression combined with craniotomy and hematoma clearance under intracranial pressure monitoring,and intracranial pressure monitoring and management were carried out after operation.The rate of bone flap acceptance during operation,incidences of complications such as re-bleeding,scalp exudation,intracranial infection and cerebral infarction after operation,rate of re-operation and Glasgow outcome scale scores 6 months after injury were compared and analyzed between the two groups.Results Five patients had midway withdrawal (2 from the control group and 3 from the treatment group),and 101 patients (49 from the control group and 52 from the treatment group) were included in the statistical analysis.The rate of bone flap acceptance in the treatment group (69.2％) was significantly higher than that in the control group (24.5％,P＜0.05).The incidences of complications such as bleeding,scalp exudation,intracranial infection and cerebral infarction (11.5％,7.7％,3.8％,and 13.5％) were significantly lower than those in the control group (30.6％,22.4％,16.3％,and 34.7％,P＜0.05).The re-operation rate (3.8％) was significantly lower than that in the control group (16.3％,P＜0.05).Good recovery rate in the treatment group (76.9％) was significantly higher than that in the control group (55.1％,P＜0.05).The mortality rate (7.7％) was significantly lower than that of the control group (22.4％,P＜0.05).Conclusion For patients with severe cerebral hemorrhage,controlled decompression under intracranial pressure monitoring combined with craniotomy and hematoma removal can significantly improve the rate of bone flap acceptance,reduce the rate of second-stage cranioplasty,reduce the incidence of complications and re-operation rate,and more effectively improve the quality of life and prognosis of patients.

Aim To investigate the antitumor and antiangiogenic effects of combined low-dose cyclophosphamide(CTX)and recombined VEGF protein vaccine.Methods In this experiment,H22 hepatocellular carcinoma model was established in BALB/c mice.Mice were randomly divided into four groups: control group,CTX group(CTX),VEGF protein vaccine group(V2)and CTX plus V2 group(CTX+V2).The anti-tumor efficacy and antiangiogenic effect were investigated using a subcutaneous tumor model and an intradermal tumor model.Western blot and ELISAwere further adopted to detect the specific anti-VEGF antibody.Results CTX+V2 group displayed a lower tumor volume and tumor weight than either the single therapy group in the subcutaneous tumor model(P<005 vs V2,P<001 vs CTX).Meanwhile,CTX+V2 was more effective for antagonizing tumor-associated angiogenesis compared with either the single therapy(P<005 vs V2,P<001 vs CTX).After CTX+V2 immunization,high titer of anti-VEGF antibody was detected by ELISA and verified by Western blot.Conclusion The therapy of CTX combined with V2 has significant synergistic effect against H22 hepatocellular carcinoma.

Objective To investigate the effects of triptolide (TP) on proliferation and apoptosis of acute myeloid leukemia cell line MV411 with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD), and its effect on PI3K-Akt-mTOR pathway. Methods MTT assay was used to detect the proliferation inhibition of MV411 cell proliferation treated by different concentrations of TP in 24, 48 and 72 h. Flow cytometry was used to measure the cell apoptosis rate at 48 and 72 h. Real-time fluorescent quantitative PCR was used to detect the expression of FLT3, PTEN, PI3K, Akt, mTOR mRNA on PI3K-Akt-mTOR pathway. Results After treated by 0, 5, 10, 20, 40, and 80 nmol/L TP in 24, 48 and 72 h, the viability of MV411 cells was inhibited in a time-dose dependence manner. MV411 cells was treated by 0, 10, and 20 nmol/L TP after 48 and 72 h, the cell apoptosis rates were rising with the increasing of drug concentration, there were statistical significances [48 h:(3.30±0.20) %, (17.10±0.36) %, (35.67±0.61) %, 72 h: (7.37±0.32) %, (49.33± 0.40)%, (68.92±0.11)%, all P=0.000]. The expressions of FLT3, PI3K, Akt, and mTOR mRNA were down-regulated and PTEN mRNA expression was up-regulated by TP. Conclusion TP may inhibit the proliferation and induce apoptosis of MV411 cells by affecting the expression of PI3K-Akt-mTOR pathway related genes.

Objective To explore the effect of proliferation and apoptosis,and research its mechanism associated with RAS/extracellular signal regulated kinase/mitogen-activated protein kinase (RAS/ ERK/MAPK) in Fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) mutation acute lymphocytic leukemia cell line MV4-11 cells treated by triptolide (TP).Methods MV4-11 cells were respectively treated by triptolide with diverse concentrations and different times.The proliferation inhibition rate was measured by methyl thiazolyl tetrazolium,the apoptosis rate was detected by flow cytometry,the mRNA expressions of FLT3,RAS,ERK,forkhead box protein M1 (FOXM1),and c-Myc were analyzed by realtime fluorescent quantitative polymerase chain reaction (PCR).Results The proliferation inhibition rate markedly increased in a dose-time dependent manner after MV4-11 cells were treated by TP.After cells were treated with 10,and 20 nmol/L TP,respectively,the apoptosis rates at 48 h were (17.30 ± 0.56) ％,(35.77 ±0.55)％,and those at 72 h were (49.83 ±0.45)％,(68.90 ±0.75)％ correspondingly.PCR data showed that the mRNA level of FLT3 mRNA decreased obviously,and that of RAS,ERK,FOXM1,and c-Myc also decreased.Conclusions Triptolide could significantly inhibit the MV4-11 cells proliferation and induce cell apoptosis,and its mechanism might be through inhibiting the expression of related genes on RAS/ERK/MAPK signaling pathway.

[Objective] This prospective study was conducted to assess the influence of overnight orthokeratology (OK) on low,middle and high myopia in southern Chinese teenagers.[Methods] Fifty-four subjects (107 eyes,age 11.65 ± 2.44 years,mean±SD)who matched the inclusion criteria for OK were enrolled in the study.The subjects were divided into three groups according to the different refraction baseline (low myopia group:0.75 ～ 3.00 D,51 eyes;middle myopia group:3.25 ～ 5.00 D,36 eyes;high myopia group:5.25 ～ 9.00 D,20 eyes).Refraction,central corneal thickness,corneal keratometry,anterior chamber depth and axial length (AL) were measured at baseline and after 1 years using ocular biometry.The changes were evaluated and compared among the groups.Results were analyzed by SPSS 17.0 software.[Results] Fifty-four subjects (total 107 eyes) completed the 1-year follow-up examinations.At baseline,the spherical equivalent refractive error (SER) was-2.15 D ± 0.63 D,-4.20 D ± 0.54 D,and-6.63 D ±1.18 D in three groups.The axial length was (24.50-± 0.66) mm,(25.16-± 0.65) mm,and (26.35 ± 0.87) mm in the three groups,respectively.There were no significant differences among the groups in initial central corneal thickness,anterior chamber depth,central,flat and steep kerotometry.The increase in axial length during the 1-year study period was (0.22 ± 0.27) mm,(0.09-± 0.22)mm and (0.02-± 0.22) mm in three groups,respectively,and the difference was significant between low myopia group and middle myopia group (P =0.04),low myopia group and high myopia group (P =0.008),whereas no significant difference was found between middle myopia group and high myopia group (P =0.35).Spherical equivalence decreased dramatically in three groups after one year (all P ＜ 0.0001).And the difference was significant between low myopia group and middle myopia group (P ＜ 0.0001),low myopia group and high myopia group (P ＜ 0.0001).Central,flat and steep kerotometry after one year became flatten in all group (all P ＜0.0001).There were no significant differences in the change of central corneal thickness,anterior chamber depth and corneal diameter.[Conclusions] OK lens is more effective in middle and high myopia control than in the low myopia control.The change in corneal keratometry may be one of the main factors influencing the OK treatment effect.