Objective:To investigate the efficacy and major adverse cardiovascular events(MACE)in elderly patients with ischemic cardiomyopathy-induced heart failure with reduced ejection fraction(HFrEF)treated with Sacubitril-valsartan at 3 years of follow-up.Methods:This was a single-center retrospective cohort study.The elderly patients with ischemic cardiomyopathy-induced HFrEF aged 60-85 years were diagnosed and treated in Beijing Anzhen Hospital from January 2018 to January 2020.A total of 120 continuously included elderly HFrEF patients treated with sacubitril valsartan were enrolled as the observation group, and 120 age-, gender-and B-type natriuretic peptide-matched elderly HFrEF patients treated with angiotensin converting enzyme inhibitor/angiotensin receptor blocker(ACEI/ARB)were enrolled as the control group.Structure and function of the heart were detected at 1-year follow-up.The MACE was observed, including cardiogenic death, readmission due to heart failure worsening and malignant arrhythmia at 3-year follow-up.Results:During follow-up(23.5±11.6)months, the visit of 18 cases was lost.There were no significant differences in the age, medical history, blood pressure between the two groups( P>0.05). Follow-up results showed that the improvement was better in observation group than in control group as follows: the left ventricular ejection fraction(45.8±9.4)% vs.(40.7±8.5)%, left ventricular end-diastolic diameter(56.5±8.3)mm vs.(59.2±7.3)mm, left ventricular end-systolic diameter(42.5±11.2)mm vs.(45.7±9.6)mm, left atrium inner diameter(49.1±8.7)mm vs.(51.2±7.7)mm, and left ventricular mass index(111.3±34.3)g/m 2vs.(119.7±31.5)g/m 2( t=4.41, 2.68, 2.38, 1.98 and 1.98, respectively, P<0.01 or 0.05). The rates of readmission due to heart failure worsening and the incidence of MACE were lower in the observation group than in the control group(21.7% or 26/120 vs.36.7% or 44/120, and 45.0% or 54/120 vs.71.7% or 86/120, χ2=6.54 and 17.55, P<0.05 or 0.01). In patients with the grade Ⅲ and Ⅳ New York Heart Association(NYHA)cardiac function, the incidence of MACE were lower in the observation group than in the control group(75.0% or 9/12 vs.100.0% or 14/14, χ2=5.10, P<0.05). Conclusions:Sacubitril-valsartan can improve cardiac structure and function, and decrease the incidence of MACE in elderly patients with HFrEF induced by ischemic cardimyopathy.

Sixty-eight cases of naevus fusco-caeruleus zygomaticus were observed from 1990 to 1996 in our clinic. In order to differentiate easily from other pigmented disorders, detailed analysis was carried out. The results showed that this condition principally occurred in young or middle-aged women, manifested as dark greyish spots scattered bilaterally and symmetrically on the zygomatic region. Histopathologically, there were elongated, slender spindle-shaped melanocytes scattered in the upper portion of the dermis, their long axes run parallel to the collagen fibres. It suggests that the naevus reported differs clinically and histopathologieally from naevus of Ota.

cases of vitreous haemorrhage secondary to Eales′ disease were selected for vitrectomy. According to the duration of vitreous haemorrhage, the patients were divided into two groups : Group 1 (20 eyes)-early vitrectomy group with a duration between 3 to 6 months; Group 2(20 eyes)-deferred vitrectomy group with a duration of more than 6 months . All the patients were followed up for a minimum period of 3 months following vitrectomy. The eyes in Group 1 showed a preoperative ultrasonic picture of complete posterior detachment, a final visual acuity of 0.6 or better in 13(65%) was achieved in mild vitreous organization on kinetic echography. Poor visual outcome in the deferred group was secondary to cystoid macular oedema, macular scar, macular pucker formation and macular degeneration. Improved visual outcome in the early vitrectomy group is probably due to haemorrhage and its products which have no time to damage the macula and to advance into macular traction and cystoid macular oedema.

Objective To investigate the serum levels of CXC, CC and C chemokines, and to esti-mate the relationship among the three kinds of chemokines as well as that between these chemokines and other Th1 and Th2-related cytokines, in patients with atopic dermatitis (AD). Methods Fifty-one patients with atopic dermatitis, including 35 males and 16 females with an average age of 17 years and disease course of 13.6 years were enrolled into this study together with 34 normal human controls. ELISA was used to measure the serum levels of monokine induced by IFN-γ (Mig), thymus and activation-regulated chemokine (TARC), cutaneous T cell-attracting chemokine (CTACK) and lymphotactin (Ltn) in these subjects. Severity of AD was assessed according to SCORAD. Results The serum levels were 79.6±28.0 ng/L for Mig, 349.9±91.5 ng/L for TARC, 747.4±359.4 ng/L for CTACK and 141.0±68.4 ng/L for Ltn in patients, significantly higher than those in the normal controls (63.8±26.5 ng/L, 219.4±82.1 ng/L, 294.3± 64.9 ng/L, 80.9±54.2 ng/L, P < 0.05, 0.01, 0.01, 0.01 respectively). A significant correlation was observed between the serum level of CTACK and SCORAD score in patients (r = 0.343, P < 0.05). Similarly, the percentage of body surface area (BSA) involved positively correlated with the serum levels of Mig, TARC, CTACK and Ltn. Furthermore, there was a significant correlation between the serum level of Ltn and TARC (r=0.444, P< 0.01) as well as CTACK (r=0.572, P< 0.01), between that of CTACK and TARC (r=0.524, P< 0.01), and between that of TARC and Mig (r=0.313, P< 0.05). Conclusion The CXC, CC and C chemokines might be involved in the pathogenesis of AD. Further more, CTACK level could serve as a good indicator for the severity of AD.

Objective To determine the effect of NS3 and NS4A proteins of Zika virus on the neuronal migration in vivo.Methods To identify the coding sequence of NS3 and NS4A,the genome of Zika SZ01 was sequenced by rapid amplification of cDNA ends (RACE) and reverse-transcription PCR,then NS3 and NS4A was constructed in pCIG vector fused with Flag-tag to express these proteins.And then these plasmids was transfected into the embryo brain of E13.5 mice by in utero electroporation,the distribution of the cells which express these proteins in the cortex was detected by Flag,eGFP and TBR1 fluorescence in E18.5 mice through immunohistochemistry so as to assess the influence of viral proteins on the neuronal migration of mouse cortex.Results 1) Sequence results showed that the amino acid sequence of NS4A is consistent with NCBI data,while NS3 has 1 amino acid mutant.2) As the fluorescence of Flag and eGFP can co-localization,the eGFP fluorescence signal marks the cells that have expressed these virus proteins in cortex.3) TBR1 fluorescence shows the distribution of the cells that express NS4A in vivo are significantly different from pCIG control and NS3 (P＜0.001).Conclusions The NS4A protein of Zika virus may affect the neuronal migration in vivo.

Objective To investigate the changes of expression of quinone oxidoreductase-1 (NQO1) and heme oxygenase-1 (HO1) at protein and mRNA levels in the brains of rats with chronic fluorosis,effect on NF-E2-related factor 2/antioxidant response element (Nrf2/ARE) signal pathway,and reveal the mechanism of brain damage induced by the disease.Methods SD rats were randomly divided to two groups of 30 each (half females and half males),e.g.the normal control group (drinking water containing less than 0.5 mg/L of fluorine) and fluoride exposed group (drinking water containing 50.0 mg/L sodium fluoride,NaF).All rats were examined at the 10 months after feeding NaF.Dental fluorosis of rats was observed; the fluoride contents in urine and bone were detected by fluoride-ion selective electrode; protein and mRNA levels of NQO1 and HO1 in brains were detected by Western blotting and quantitative real timePCR,respectively.Results The dental fluorosis was observed,and contents of fluoride in urine [(2.16 ± 0.39)mg/L] and bone [(211.07 ± 40.52)mg/kg] determined in the rats of the fluoride group were higher than those of controls [(1.70 ± 0.24)mg/L,(34.67 ± 11.15)mg/kg,t =2.11,3.23,all P＜ 0.05].The protein expression levels of NQO1 and HO1 in the brains of rats with fluorosis [(255.2 ± 14.3) ％ and (187.2 ± 11.1)％] were also higher than those of controls [(100.0 ± 12.2)％,(100.0 t 8.9)％,t =2.14,2.05,all P ＜ 0.05]; the mRNA levels of NQO1 and HO1 [(210.2 ± 9.8)％ and (154.5 ± 7.4) ％] in the rats of the fluoride group were increased as compared to those of controls [(100.0 ± 10.4)％,(100.0 ± 9.7)％,t =2.33,2.75,all P ＜ 0.05].Conclusion The expression of NQO1 and HO1 in brain of rats with fluorosis are significantly increased,which may be due to the activation of Nrf2/ARE signal pathway and may play a compensative role in enhancing antioxidant ability.

Objective To detect the expression of muscarinic acetylcholine receptors (mAChRs) at mRNA and protein levels in the brain of rats with chronic fluorosis and to reveal the role of the receptors in brain injury and learning and memory deficits.Methods Sixty healthy SD rats were divided into two groups (30 rats in each group,half males and half females) by random number table method according to body weight.In the control group,the rats were fed with drinking water containing no more than 0.5 mg/L fluoride; in the fluoride group,the rats were fed with high doses of sodium fluoride in drinking water (50.0 mg/L).Each group was fed with normal diet (6.2 mg/kg).After being exposed to fluoride for 10 months,behavioral performance was measured with Morris water maze,including the escape latency time and the numbers of crossing platforms.After being sacrificed,rat brains were taken and weighted.M1 and M3 subunits at mRNA and protein levels were detected by real-time PCR and Western blotting,respectively; the correlation between protein levels of the receptor subunits and the ability of learning and memory was analyzed.Results In fluoride group,the escape latency time [(21.68 ± 2.90)s] was significantly longer than that of control group [(6.14 ± 1.71)s,t =0.289,P ＜ 0.05]; and the number of crossing platforms [(11.62 ± 2.26)times] was significantly decreased as compared to that of control group [(19.00 ± 3.69)times,t =0.352,P ＜ 0.05].Furthermore,the mRNA expression [(17.07 ± 6.89)％,(12.25 ± 5.03)％] and the protein levels [(71.07 ± 6.89)％,(32.25 ± 4.66)％] of M1 and M3 receptors in rat brains were significantly lower as compared to those of controls [(100.00 ± 3.00)％,(100.00 ± 2.15)％ and (100.00 ± 9.01)％,(100.00 ± 10.33)％,t =0.210,0.157,0.095,0.296,all P ＜ 0.05].The escape latency and M1,M3 protein levels were negatively correlated (r =-0.683,-0.700,all P ＜0.05),and the number of space exploration and M1,M3 protein levels were positively correlated (r =0.867,0.837,all P ＜ 0.05).Conclusion Declined expression of mAChRs at mRNA and protein levels have been detected in the brain of rats with chronic fluorosis,which may be one of the main mechanism concerning the learning and memory deficits.

Objective To observe the changes of learning and memory ability and detect the expression of muscarinic acetylcholine receptor (mAChR,M receptor) at mRNA and protein levels in brains of offspring rats with chronic fluorosis,and to reveal the mechanism of the central nervous system damage.Methods Forty healthy SD rats were divided into two groups (20 in each group,half male and half female) by random number table according to body weight.In the control group,the rats were fed with drinking water containing no more than 0.5 mg/L fluoride;in the fluoride group,the rats were fed with high dose of sodium fluoride in drinking water (50.0 mg/L fluoride).Each group was fed with normal diet (6.2 mg/kg fluoride).After exposed to fluoride for 6 months,each group was mated,and brains of newborn offspring rats aged 1,7,14,21 and 28 days were taken,and expression of M1 and M3 receptors at mRNA and protein levels were analyzed by real-time PCR and Western blotting,respectively.Behavioral changes were measured by Morris water maze test at the 28 days after birth.The correlations between protein levels of M1 and M3 receptors and the ability of learning and memory at the 28 days after birth were analyzed.Results In fluoride group of the offspring rats at 28 days after birth,the escape latency time [(35.61 ± 9.00)s] was significantly longer than that in control group [(8.46 ± 3.09)s,P ＜ 0.05],while the numbers of crossing the platforms and the time of staying the platforms [(5.00 ± 2.90)times,(16.66 ± 2.79)s] were significantly decreased as compared to that of control group [(15.17 ± 3.66)times,(22.51 ± 2.66)s,all P ＜ 0.05].Furthermore,the mRNA expression and the protein levels of M1 and M3 receptors in rat brain at each phase in fluoride group were significantly decreased as compared to controls [M1 mRNA in control groups:(100.00 ± 11.00)％,(100.00 ± 17.57)％,(100.00 ± 9.14)％,(100.00 ± 7.52)％,(100.00 ± 15.78)％;M1 mRNA in fluoride groups:(20.47 ± 8.07)％,(14.00 ± 4.53)％,(16.57 ± 7.62)％,(25.56 ± 12.78)％,(16.27 ± 4.82)％;M3 mRNA in control groups:(100.00 ± 16.30)％,(100.00 ± 14.40)％,(100.00 ± 7.20)％,(100.00 ± 14.31)％,(100.00 ± 13.16)％;M3 mRNA in fluoride groups:(29.17 ± 8.00)％,(12.77 ± 2.22)％,(26.40 ± 7.20)％,(15.74 ± 3.55)％,(28.14 ± 7.53)％;M1 protein in control groups:(100.00 ± 2.24)％,(100.00 ± 8.30)％,(100.00 ± 4.61)％,(100.00 ± 13.78)％,(100.00 ± 11.72)％;M1 protein in fluoride groups:(20.47 ± 8.07)％,(14.00 ± 4.53)％,(16.57 ± 7.62)％,(25.56 ± 12.78)％,(16.27 ± 4.82)％;M3 protein in control groups:(100.00 ± 16.30)％,(100.00 ± 14.40)％,(100.00 ± 7.20)％,(100.00 ± 14.31)％,(100.00 ± 13.16)％;M3 protein in fluoride groups:(29.17 ± 8.00)％,(12.77 ± 2.22)％,(26.40 ± 7.20)％,(15.74 ± 3.55)％,(28.14 ± 7.53)％,P ＜ 0.05 or ＜ 0.01].The escape latency and M1,M3 receptors protein levels were negatively correlated (r =-0.827,-0.742,all P ＜ 0.05),and the number of space exploration and M1,M3 receptors protein levels were positively correlated (r =0.843,0.806,all P ＜ 0.05).Conclusion The expression of M receptor at protein and mRNA levels in offspring rat brains of different ages are significantly declined,which might be one of the mechanism of the decreased ability of learning and memory induced by fluoride toxicity.

Objective To observe the expression of NF-E2-related factor 2 (Nrf2) and Kelch-like ECH-associated protein-1 (Keap1) at mRNA and protein levels in the brain of rats with chronic fluorosis and to reveal the mechanism of brain damage induced by the factors.Methods Ninety SD rats were divided into three groups (30 rats in each group,half male half female) by the random number table method according to body weight.The control group was fed with normal tap-water,high-fluoride group with 50 mg/L fluoride (NaF) added in drinking water;and the high-fluoride plus vitamin E (Vit E) group with the same dose of NaF as the high-fluoride group,but giving 5 mg/kg Vit E by intragastric administration.The experiment period was 10 months.The fluoride contents in urine and bone were detected by fluoride-ion selective electrode.The protein and mRNA levels of Nrf2 and Keap1 in brain of rats were detected by Western blotting and quantitative real time PCR,respectively.The activity of superoxid dismutas (SOD) and the content of lipid peroxidation (MDA) were measured by biochemistry methods.Results Dental fluorosis was detected in high-fluoride group.The differences of fluoride contents in urine and bone were statistically significant between groups (F =6.87,182.87,all P ＜ 0.05).The urine fluoride [(2.16 ± 0.39),(2.07 ± 0.15)mg/L] and bone fluoride [(211.07 ± 40.52),(82.09 ± 28.60)mg/kg] in the high-fluoride and high-fluoride plus Vit E groups were higher than those of the control group [(1.70 ± 0.24)mg/L,(34.67 ± 11.15)mg/kg,all P ＜ 0.05].The differences of mRNA and protein levels of Nrf2 and Keap1 in brains of rats,SOD activity,MDA content were statistically significant between groups (F =654.33,432.87,447.45,398.88,68.34,68.34,all P ＜ 0.05).The mRNA levels of Nrf2 and Keap1 [(320.18 ± 6.83)％,(267.37 t 7.22)％] were increased compared to those of control group [(100.00 ±3.00)％,(100.00 ± 2.75)％,all P ＜ 0.05];the protein levels of Nrf2 and Keap1 [(283.28 ± 6.89)％,(196.32 ± 5.57)％]were also raised compared with those of control group [(100.00 ± 8.71)％,(100.00 ± 9.23)％,all P ＜ 0.05];the activity of SOD [(22.10 ± 2.10)μ,mol/kg] in brain of rats in fluoride group was significantly lower and the content of MDA [(8.63 ± 0.77) μmol/kg] was higher than those of control group [(35.05 ± 2.98),(1.25 ± 0.64) μmol/kg,all P ＜ 0.05].In high-fluoride plus Vit E group,the mRNA levels of Nrf2 and Keap1 [(243.23 ± 5.34)％,(180.54 ± 4.48)％] and the protein levels of Nrf2 and Keap1 [(210.88 ± 4.79)％,(150.68 ± 6.49)％] were lower than those of high-fluoride group (all P ＜ 0.05);the activity of SOD [(26.33 ± 1.84)μmol/kg] was significantly higher and the content of MDA [(4.88 ± 0.84)μmol/kg] was lower than that of high-fluoride group (all P ＜ 0.05).Correlation analysis showed that increased levels of Nrf2 and Keap1 were positively correlated with the level of MDA in rat brain (r =0.69,0.33,all P ＜ 0.05),but negatively correlated with the activity of SOD (r =-0.78,-0.80,all P ＜0.05).Conclusion The expression of Nrf2 and Keap1 in brain of rats with fluorosis is significantly increased and positively correlated with the content of fluoride in bone and the level of oxidative stress,whereas vit E can attenuate these abnormal changes induced by fluoride,which might be one of the mechanisms of brain damage of the disease.

Objective To report a consanguineous family with lamellar ichthyosis and to detect the mutations in transglutaminase 1 (TGM1) gene in this family. Methods Genomic DNA was extracted from the blood samples of a 19-year-old male patient with lamellar ichthyosis, his family members and 100 normal human controls. PCR was carried out to amplify all the encoding sequences (15 exons) and adjacent flanking sequences of TGM1 gene followed by bidirectional sequencing. Results A C1666T mutation in the 11th exon in TGM1 gene, which resulted in the substitution of ACA (threonine) by ATA (isoleucine) at codon 529, was detected in the proband, while both his parents carried the C1666T mutation in heterozygous form, and his sister was a C/C homozygote. None of the 100 normal control individuals carried the mutation in TGMlgene. Conclusions The de novo mutation from ACA (threonine) to ATA (isoleucine) at codon 529, may contribute to the development of lamellar ichthyosis. Consanguineous marriage can increase the risk for lamellar ichthyosis by raising the probability of homozygosis of C 1666T mutation in TGM 1 gene.