Objective To investigate the effect of adenovirus-mediated IL-24 (Ad-IL-24)gene expression on the apoptosis in a human squamous cell carcinoma cell line COLO 16, and to explore the underlying molecular mechanism. Methods Cultured COLO 16 cells were divided into two groups to be transfected with an adenovirus vector carrying the IL-24 gene (Ad-IL-24 group)or green fluorescent protein (Ad-GFP group), while those receiving no treatment served as the control group. After culture for different durations, qPCR was performed to quantify IL-24 gene expression, methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of COLO 16 cells, flow cytometry to detect the apoptosis of COLO 16 cells, laser scanning confocal microscopy (LSCM) to observe the morphological changes of COLO 16 cells, Western blot to determine the levels of Bax and Bcl-2 proteins and to evaluate the activation of caspase-3, qPCR to determine the levels of Bax and Bcl-2 mRNAs, an immunofluorescence assay to observe the expression of Bax and Bcl-2 proteins. Statistical analysis was carried out by a two-sample t-test with the SPSS 19.0 software. Results MTT assay showed that the proliferation of COLO 16 cells in the Ad-IL-24 group was significantly inhibited as early as 4 days after the transfection; thereafter, the inhibitory effect increased in a time-dependent manner, and peaked on day 6(P<0.05). However, there was no significant difference in cellular proliferative activity between the Ad-GFP group and control group (P>0.05). Flow cytometry revealed that the apoptosis rate was significantly higher in the Ad-IL-24 group(13.10%± 0.92%)than in the control group(3.69%± 0.36%, P<0.05)and Ad-GFP group(3.39%± 1.06%, P<0.05), but no significant difference was observed between the control group and Ad-GFP group (P>0.05). LSCM demonstrated that the apoptosis of COLO 16 cells was accelerated in the Ad-IL-24 group. The immunofluorescence assay, Western blot and qPCR all showed that the mRNA and protein expressions of Bax were increased, but those of Bcl-2 were decreased in the Ad-IL-24 group compared with the Ad-GFP group and control group. Moreover, Western blot showed a protein band that could specifically bind to the anti-cleaved caspase-3 antibody in the Ad-IL-24 group, but not in the Ad-GFP group or control group. Conclusions Ad-IL-24 can induce apoptosis in human COLO 16 squamous cell carcinoma cells, probably by up-regulating Bax expression, down-regulating Bcl-2 expression, and activating caspase 3.

BACKGROUND: At present, treatment of diabetes mellitus with western medicine starts from the angle of stimulating injured β cell of islet to strength insulin secreting. Treatment with traditional Chinese medicine is very important in protecting injured β cells.OBJECTIVE: To observe the protective effect of lycium barbarum polysaccharide and nourishing yin and activating blood compound recipe on pancreatic islet function in rats with streptozotocin-induced diabetes mellitus.DESIGN: A randomized and controlled animal experiments.SETTING: First Affiliated Hospital and Medical College of Henan University of Science and Technology.MATERIALS: Totally 70 male Wistar rats, aged 6 weeks, with body mass of 200 g, were used in this experiment.METHODS: This experiment was carried out at the Animal Room of Henan University of Science and Technology from October 2001 to April 2002. Totally 70 rats were divided into two groups: normal control group (n=10) and modeling group (n=60). Totally 30 successful model rats were randomly subdivided into 3 groups: streptozotocin model group, compound recipe treated group and lycium barbarum polysaccharide-treated group,with 10 rats in each group ;Intragastric administration of 60 g/kg Chinese herb compound recipe for nourishing yin and activating blood was performed on the rats in the compound recipe treated group at 9:00 every day (huangqi, shanyao, gegen respectively 30 g, gouqizi, danshen, huafen,digupi respectively 15 g, danggui, zhimu respectively 12 g, danpi, wuweizi respectively 10 g, shuizhi 5 g, shanyurou 20 g, etc, provided by drug store of First Hospital Affiliated to Henan University of Science and Technology,were decocted and condensed to be equivalent crude drug 3.0 kg/L with routine method). Intragastric administration of 0.5 g/kg lycium barbarum polysaccharide was given in the lyciun barbarum polysaccharide treated group (lycium barbarum polysaccharide was isolated and purified from ningxia barbary wolfberry fruit). Intragastric administration of comparative volume of normal saline was performed in the streptozotocin model group and normal control group, totally 3 weeks. The levels of fasting blood glucose (FBG) and insulin were measured at 3 weeks before and after experiment, and function index of β cell of islet was calculated; Pancreas was taken out to detect SOD activity, NOS activity and concentrations of NO and MDA were also detected.RESULTS: Ten rats in normal control group and 30 successful model rats in model group entered the stage of result analysis. ① In the initial stage of experiment, FBG level in each modeling group was significantly increased (t=16.51 to 10.07,P ＜ 0.01), while fasting insulin level and function index of β cell were significantly decreased (t=13.64 to 100.99 ,P ＜ 0.01 ) in comparison with normal control group. ② At 3 weeks after experiment, levels of FBG, NO , MDA , NOS activity were significantly decreased in the compound recipe treated group and lycium barbarum polysaccharide group compared with streptozotocin model group (t=8.08 to 72.68 ,P ＜ 0.01 ), while level of fasting insulin and function index of β cell, SOD activity were significantly increased in the compound recipe treated group and lycium barbarum polysaccharide group compared with streptozotocin model group (t=4.39 to 17.87,P ＜ 0.05-0.01 ). ③ Levels of FBG and MDA , NOS activity and concentration of NO were significantly decreased in the compound recipe treated group, and fasting insulin and function index of βcells were significantly increased in comparison with lycium barbarum polysaccharide group(P ＜ 0.01 ).CONCLUSION: Lycium barbarum polysaccharide and nourishing yin and activating blood compound recipe increase the level of fasting insulin and function index of βcell of rats with streptozotocin induced diabetes mellitus, decrease the level of blood glucose, improve the function of βcell of islet, which might be related to the increase of pancreatic islet SOD activity and decrease NOS activity.

Objective To study the effect of abnormal left ventricular diastolic function(LVDF)on the onset and severity of atrial or ventricular arrhythmia in elderly essential hypertensive patients.Methods The 210 elderly essential hypertensive patients were enrolled in this study. Their arrhythmias were monitored by 24-hour ambulatory electrocardiogram. The essential hypertensive patients were referred for Doppler echocardiography to evaluate left ventricular function, while patients with abnormal systolic function were excluded, and then the patients were classified as normal LVDF and abnormal LVDF including, impaired relaxation, pseudonormal, and restrictivelike filling patterns. Results In 210 elderly essential hypertensive patients, 147 (70%) cases were diagnosed as atrial arrhythmia and 102 (49%) cases as ventricular arrhythmia (χ2 = 19. 975, P ＜ 0.05 ).Morbidities of atrial (89%) or complex atrial arrhythmia (49%) as well as ventricular (63%) orcomplex ventricular arrhythmia (30%) were significantly higher in abnormal LVDN group than in normal LVDN group (40%, 13%, 26% and 7%, χ2 = 56. 723 、 28. 359 、 28. 076、15. 9102 , all P＜0. 05). The morbidities of arrhythmias were higher in hypertensive patients with pseudonormal and restrictiveike filling patterns than in other groups 93.6% and 96. 4%. Conclusions Abnormal left ventricular diastolic function affects on the onset and severity of atrial or ventricular arrhythmia in elderly essential hypertensive patients, and complex atrial or ventricular arrhythmia is easier found in hypertensive patients with pseudonormal and restrictivelike filling patterns.

Objective To study the effects of paeonol on the tyrosinase activity and melanogenesis in co-culture model of human melanocytes and keratinocytes. Methods Melanocytes and the co-culture model of human melanocytes and keratinocytes were cultivated and the proliferation of melanocytes and the co-cultures was measured by MTT colorimetric assay. The tyrosinase activity and melanin level were measured by enzymic method. Results The melanin synthesis and tyrosinase activity were markedly suppressed by paeonol in a dose-dependent manner at the concentrations of 50?mol/L, 100?mol/L, and 200?mol/L in both melanocytes and co-cultures. The significant stronger suppression was observed with 100?mol/L and 200?mol/L of paeonol than that with controls (P

Objective To evaluate the inhibitory action of ethanolic extracts from196kinds of Chi-nese herbs on tyrosinase.Methods Tyrosinase inhibitory activity was determined by the dopachrome method using L-DOPA as the substrate and the amount of dopachrome in the reaction mixture was measured by spec-trophotometer.Results Nine Chinese crude extracts[Glycyrrhiza glabra L.,Melaphis chinensis(Bell.)Baker,Cryptotympana atrata Fabricius,Paeonia suffruticosa Andr.,Sophora flavescens Ait.,Spirea thunbegii Sieb.et Bl.,Xanthium sibiricum Patr.,Morus alba L.,Rheum palmatum L.]showed potent inhibitory action on tyrosi-nase compared with positive control arbutin(1mmol/L,P0.05).Conclusion The results suggest that20kinds of crude extracts from Chinese herbs[Glycyrrhiza glabra L.,Melaphis chinensis(Bell.)Baker,Cryptotympana a-trata Fabricius,Paeonia suffruticosa Andr.,Sophora flavescens Ait.,Spirea thunbegii Sieb.et Bl.,Xanthium sibiricum Patr.,Morus alba L.,Rheum palmatum L.,Artemisia anomala S.Moore,Hemistepta lyrata Bunge,Lycium chinensis Mill.,Dipsacus asper wall.,Gastrodia elata Bl.,Forsythia suspensa(Thunb.)Vahl.,Acer gin-nala Maxim.,Glycyrrhiza uralensis Fisch,Patrinia sabiosaefolia Fisch.,Buxus sinica(Rehd.et Wils.)Ching,Lonicera japonica Thunb.,]inhibit tyrosinase and may be used as depigmentaty agents for pigmentary skin dis-eases caused by abnormal tyrosinase activity.

Objective To estimate the effect of a tretinoin derivative ECPIRM on retinoic acid receptors (RARs),and to observe skin irritation responses to it in mice.Methods Cultured SCL-1 cells were divided into 2 groups to be treated with culture medium containing 10 μmol/L ECPIRM (ECPIRM group) or 10 μmol/L all-trans retinoic acid (ATRA) (ATRA group) for 24 hours,and those treated with drug-free culture medium served as the control group.Western blot analysis and real-time fluorescence-based quantitative PCR were performed to quantify the protein and mRNA expressions of RARs (RARα,RARβ,RARγ and RXRα) respectively.In addition,real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expressions of two target genes of the activated RAR signaling pathway,i.e.,cytochrome P450 26A1 (CYP26A1) and tazarotene-induced gene 1 (TIG1).Eight BALB/c mice were equally divided into 2 groups to be topically treated with 0.075％ ECPIRM gel or 0.05％ ATRA cream at equal molar concentrations on the shaved skin once daily for 21 successive days.Skin irritation reactions were assessed in these mice.Results Compared with the control group,the ATRA group showed significantly increased protein and mRNA expressions of RARα,RARβ and RARγ (all P ＜ 0.05).The mRNA expressions of CYP26A1 and TIG1 genes in the ATRA group were 25.49 and 3.88 times that in the control group respectively (both P ＜ 0.01).However,there was no significant difference in the protein expressions of RARα,RARβ,RARγ and RXRα,or mRNA expressions of RARα,RARβ,RARγ CYP26A1 and TIG1 between the ECPIRM group and control group (all P ＞ 0.05).Obvious Skin irritation reactions such as erythema and desquamation were observed in BALB/c mice after 2-day topical treatment with ATRA cream,and their degree peaked after 5-day treatment.However,neither erythema nor desquamation was observed in BALB/c mice during 21-day treatment with 0.075％ ECPIRM gel.Conclusion Unlike ATRA,ECPIRM cannot activate the canonical RAR signaling pathway or cause skin irritation reactions.

Objective To analyse the changes of cerebral cortical thickness and explore the connectivity of cortical thickness and the clinical syndrome of concomitant strabismus using high-resolution magnetic resonance imaging.Methods Participants were 26 children with primary strabismus and 28 healthy children strictly matched for sex, age, education and socioeconomic level.By using Freesurfer software, the whole-brain-based analysis was perfomrmed to compare the cortical thickness between the two groups.Results Compared with the healthy control group the children with strabismus were found that the cerebral cortex became thinner, including these brain regions related with stereovision,advanced visual-attention,attention and executive control function,as well as brain areas in insula and cingulate circuit.Conclusion Concomitant strabismus in children are not only caused by pathological changes of extraocular muscles,but anatomical changes of the central nervous system, especially the brain areas involved in the advanced visual processing related with visual-motor and visual-attention, and the advanced cognitive brain regions associated with attention and executive control, as well as the anatomical changes in insula and cingulate cortex circuit, which can explain the clinical syndrome of children with concomitant strabismus.

Objective To investigate the clinical efficacy and safety of transcatheter arterial chemoemblization (TACE) using raltitrexed and lobaplatin in treating advanced hepatocellular carcinoma (HCC).Methods From March 2009 to November 2014,95 cases were treated by raltitrexed combined with lobaplatin (raltitrexed group) through TACE and 124 cases by fluorouracil combined with oxaliplatin (fluorouracil group) through TACE.Disease control rate (DCR),median progression-free survival (mPFS) time and median overall survival (mOS) time were compared between the two groups.Survival rate were analyzed using Kaplan-Meier method and Log-rank analysis in SPSS 16.0.Results The disease control rate of raltitrexed group was 91.6％ (87/95),compared with fluorouracil group of 84.6％ (105/124) in fluorouracil group (x2 =2.505,P =0.474).The mPFS of raltitrexed group was 6.8 months and that of fluorouracil group was 5.9 months (x2 =5.542,P =0.019);mOS of raltitrexed group was 13.6 months and fluorouracil group was 11.4 months (x2 =5.953,P =0.015).The main adverse reactions in the two groups were not statistically significant (P ＞ 0.05).Conclusions TACE using rahitrexed and oxaliplatin prolongs the progression free survival and overall survival time of patients with advanced hepatic carcinoma.

Objective:To evaluate the inhibitory effect of a retinoid derivative ECPIRM on proliferation of a cutaneous T-cell lymphoma (CTCL) cell line HH, and to explore its potential mechanisms.Methods:Cultured HH cells were treated with ECPIRM at different concentrations of 0 (control group) , 5, 10 and 20 μmol/L separately for 72 hours, cell counting kit-8 (CCK8) assay was performed to evaluate the effect of ECPIRM on the proliferative activity of HH cells, and flow cytometry to investigate the effect of ECPIRM on apoptosis of HH cells. Some HH cells were treated with 10 μmol/L ECPIRM for 72 hours, transcriptome sequencing was performed to investigate gene expression changes triggered by ECPIRM in HH cells, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis and gene ontology (GO) enrichment analysis were then performed to analyze differentially expressed genes in HH cells induced by ECPIRM. Reverse transcription-qPCR was subsequently conducted to verify changes in key gene expression in related pathways. Intergroup differences were analyzed by using one-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:CCK8 assay showed that the 50% inhibitory concentration (IC50) of ECPIRM on HH cells was 4.91 ± 2.48 μmol/L, the viability of HH cells significantly differed among the control group, and 5-, 10-and 20-μmol/L ECPIRM groups (100.00% ± 2.87%, 49.58% ± 4.53%, 48.36% ± 2.88%, 31.44% ± 2.46%, respectively, F=162.86, P < 0.001) , and was significantly lower in the 5-, 10-and 20-μmol/L ECPIRM groups than in the control group ( t=15.36, 15.73, 20.89, respectively, all P < 0.001) . Flow cytometry showed that there was a significant difference in the apoptosis rate among the 4 groups (11.51% ± 1.84%, 23.83% ± 5.72%, 36.19% ± 8.33%, 49.75% ± 4.10%, respectively, F=17.62, P < 0.001) , and the 10-and 20-μmol/L groups showed significantly increased apoptosis rates compared with the control group ( t=4.46, 6.92 respectively, both P < 0.01) . Transcriptomics analysis revealed that the inhibitory effect of ECPIRM on the cellular proliferative activity may be related to the metabolic regulation of steroids. As reverse transcription-qPCR revealed, the 10-μmol/L ECPIRM group showed significantly decreased mRNA expression of L-amino acid oxidase (IL4I1) , acetyl-coenzyme A acetyltransferase 2 (ACAT2) , 3-hydroxy-3-methylglutaryl-coenzyme A synthase 1 (HMGCS1) , mevalonate diphosphate decarboxylase (MVD) , 3-β-hydroxysteroid-8,7-isomerase (EBP) , very low-density lipoprotein receptor (VLDLR) , 3-hydroxy 3-methylglutaryl-CoA reductase (HMGCR) compared with the control group (all P < 0.05) . Conclusion:The retinoid derivative ECPIRM exerted marked anti-proliferative and apoptosis-inducing effects on HH cells, which may be related to the decreased expression of key genes involved in steroid metabolism.

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.