OBJECTIVE: To observe the release mode and the amount of retention in skin of podophyllotoxin liposomes in topical application. METHODS: Two kinds of podophyllotoxin liposome suspension (DPPC liposome and bean lecithin liposome) and podophyllotoxin tincture in same concentration were topically applied on the skin of porkets and the amounts of retention of drug in skin of different preparations were detected with confocal laser scanning microscope.RESULTS: The quantity of retention of podophyllotoxin DPPC liposome in epidermis and superficial layer of dermis was larger than those of other two preparations and the drug concentration in applying DPPC liposome kept higher for 48 hours.CONCLUSION:Podophyllotoxin liposome coated by DPPC is much more targetable to skin and is a fairly good topical preparation for applying on skin.

Nine compounds were isolated from the n-butanol fraction of 95% ethanol extract of the fruit of Alpinia oxyphylla Miq. with a combination of various chromatographic approaches, including MDS resin, silica gel, reverse phase C18 and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as (1R, 4R, 10R)-1β, 4α-dihydroxy-11, 12, 13-trinor-5, 6-eudesmen-7-one (1), 1β, 4β-dihydroxy-11, 12, 13-trinor-8, 9-eudesmen-7-one (2), oxyphyllenone A (3), oxyphyllenone B (4), rhamnocitrin (5), staphylionoside D (6), benzyl-1-O-β-D-glucopyranoside (7), 2-O-β-D-glucopyranosyl-(1S)-phenylethylene glycol (8), and (S)-1-phenylethyl-β-D-glucopyranoside (9). Among them, compound 1 is a new sesquiterpene, named as oxyphyllenone C; compounds 8 and 9 are new natural products; compounds 2 and 6 were isolated from the genus Alpinia for the first time, and compound 7 was isolated from A. oxyphylla for the first time.

Objective To investigate the application of preoperative autologous blood donation (PABD) in selective operation.Methods Retrospective investigations and analysis were carried out in clinical datas of 1 026 patients of PABD in selective operation from January 2016 to may 2017,comparing with those who had not performed PABD in the same time.Results ①The surgeries of PABD were mainly neurosurgery,urology,orthopedics,gynecology and hepatobiliary surgery,respectively,which accounted for 5.97％,4.90％,3.78％,3.76％ and 3.55％ of the patients in the same period;②The rate of transfusion of allogeneic red blood cells of selective operation in PABD group of neurosurgery,urology,orthopedics,gynecology and hepatobiliary surgery were 5.66％,2.51％,4.89％,0 and 4.88％,respetively,while not performed PABD group were 25.46％,28.58％,24.77％,10.62％ and 17.54％,respetively;③The rate of transfusion of the group of hemoglobin (Hb)＜ 120 g/L is significantly higher than those Hb is (120-129.9) g/L,(130-139.9)g/L and ≥ 140 g/L(7.65％vs 2.44％,3.66％ and 2.70％,P＞0.05).Conclusion ①PABD is mainly carried out in neurosurgery,urology,orthopedics,gynecology and hepatobiliary surgery;②The group of Hb＜ 120 g/L has a high rate transfusion of allogeneic erythrocyte in selective operations;③Only the PABD is improved can patients make benefit from it.

Objectives To provide genetic counseling and prenatal molecular diagnosis for two families with megalencephalic leukoencephalopathy with subcortical cysts (MLC).Methods Two MLC patients (probands 1 and 2) were admitted to the Department of Pediatrics of Peking University First Hospital in June 2011 and June 2009,respectively.Peripheral blood was collected and DNA sequencing was performed for genetic analysis for the two MLC patients and their parents.Amniotic fluid and villus of two fetuses (fetus 1 and 2) were collected at 21+4 and 12+3 weeks of gestational age from their mothers when they were pregnant again.The genomic DNA of the two fetuses was extracted and corresponding sites of MLC1 gene were sequenced.Haplotype analysis using a combination of 3 microsatellite markers (AR,DXS6807 and DXS6797) on chromosome X and sex determining region of Y chromosome was performed to detect maternal cell contamination.Verification of the prenatal molecular diagnosis and follow up study after birth were conducted for both fetuses.Results Macrocephaly,motor development delay and typical findings on brain MRI were identified in the two probands,and were clinically diagnosed with MLC.Compound heterozygous mutations were detected in proband 1 [c.353C＞T (p.T118M) and c.803C＞G (p.T268R)] and proband 2 [c.353C＞T (p.T118M) and c.836T＞C(p.L279P)],respectively.MLC was genetically diagnosed.Heterozygous variation in c.353[c.353C＞T (p.T118M)] and wild c.803C were identified in fetus 1,and both wild c.353C and c.836T were found in fetus 2.No maternal cell contamination was detected in both fetuses.Sequencing the corresponding sites after birth confirmed the prenatal diagnosis,and the head circumference and motor development were normal in fetus 1 at 5 months old.No macrocephaly was found and no DNA sequencing was done in fetus 2 at one month old.Conclusions Genetic counseling and prenatal molecular diagnosis for MLC families combined with clinical and genetic diagnosis are important in preventing MLC.Haplotype analysis with a combination of three microsatellite markers on chromosome X and sex determining region of Y chromosome is useful in detecting maternal cell contamination and avoiding its influence on prenatal diagnosis,and confirming the reliability of prenatal diagnosis.

The 17α-hydroxylase/17, 20-lyase deficiency (17-OHD) is a rare disease. The clinical characteristics and gene mutation of 2 late-diagnosed 17-OHD patients with testicular tumor admitted to our hospital from March 2018 to February 2019 were analyzed retrospectively. The two 17-OHD patients were female (46, XY). Laparoscopic abdominal exploration found undeveloped testicles in grey-yellow or grey-red in the groin and iliac fossa. The testicles were removed and showed malignancy in pathology study. Sequencing of the CYP17A1 gene identified c.1247G>A/c.1427T>C and c.985_987delTACinsAA/c.1306G>A complex heterozygous mutations. Taking together, the possibility of 17-OHD should be considered in patients with hypertension, hypokalemia, adrenal adenomatoid hyperplasia together with 46, XY gonadal dysplasia, so as to make early diagnosis and treatment, and avoid dysplastic testicular turning to malignancy.

Abstract The COVID-19 pandemic has posed a series of complex challenges. COVID-19 in children and adolescents is generally less severe than in adults and the elderly; however, some children and adolescents may experience severe complications and adverse health effects even after mild or asymptomatic COVID-19 infections. The article focuses on gathering the epidemic characteristics, health impact, risk factors, prevention and control measures, and vaccination status of children and adolescents with COVID-19 infection to provide recommendations for protecting children and adolescents in the post COVID-19 era.

To investigate the clinical and genetic characteristics of patients with idiopathic hypogonadotropic hypogonadism (IHH), the clinical data of 23 patients with IHH were retrospectively analyzed. Gene analyses were accomplished with whole-exome sequencing (WES) and Sanger sequencing. Functional prediction of mutation sites was conducted using two bioinformatics platforms, SIFT and Polyphen. Among the 23 patients with IHH, 9 patients carried prokinin 2 (PROKR2) gene mutations including 4 missense mutations (p.W178S, p.Y113H, p.A103V, p.R164Q), and 1 frameshift mutation (p.D42delinsDED), the remaining 14 cases were found negative in gene sequencing. Functional prediction showed that the above mutations may affect protein function suggestive of a pathogenic role of PROKR2 mutation in the patients. There were no significant differences in the levels of follicle-stimulating hormone, luteinizing hormone, testosterone, and estradiol between the IHH patients with PROKR2 gene mutation and those without. PROKR2 gene mutation might associated with IHH, and the mutations reported in the present study could enrich the pathogenic spectrum of genes.

Objective:To determine clinical curative effects of ozone therapy for pemphigus vulgaris.Methods:Ozone hydrotherapy was used as an aid treatment for 32 patients with pemphigus vulgaris.The hydropathic compression of potassium permanganate solution for 34 patients with pemphigus vulgaris served as a control.The main treatment for both groups were glucocorticoids and immune inhibitors.The lesions of patients,bacterial infection,usage of antibiotics,patient's satisfaction,and clinical curative effect were evaluated in the 2 groups.Results:There was no significant difference in the curative effect and the average length of staying at hospital between the 2 groups (P＞0.05).But rate for the usage of antibiotics was significantly reduced in the group of ozone hydrotherapy (P=0.039).The patients were more satisfied in using ozone hydrotherapy than the potassium permanganate solution after 7-day therapy (P＞0.05).Conclusion:Ozone hydrotherapy is a safe and effective aid method for pemphigus vulgaris.It can reduce the usage of antibiotics.

【Objective】 To investigate the gene frequency and polymorphism of RBC blood group systems in RhD negtive population in Hunan, so as to lay a foundation for clinical blood transfusion and construction of multiple rare blood group database. 【Methods】 Blood samples were taken from 300 RhD negative blood donors, confirmed by serological method, from June 2019 to June 2020,. RHD genotyping was performed by SSP-PCR. For blood donors with typing results as RhD negative plus RHD gene deletion, antigens genotyping of MNS, Duffy, Kell, Domrock, Diego, Kidd, Sciawnna, Colton, Lutheran and Yt RBC blood group systems were performed by SSP-PCR and analyzed by the chi square test of SPSS 20 statistical software. 【Results】 RHD gene deletions accounted for 58.67% (176 / 300) of serological D negative blood donors. The gene frequencies were as follows: MNS: GYPB^*S=0.045 5(8/176), GYPB^*s=0.954 5(168/176), GYP^*Dane=0.039 8(7/176); Duffy: FY^*A =0.965 6(170/176), FY^*B=0.034 1(6/176); Dombrock: DO^*A=0.082 4(14.5/176), DO^*B=0.917 6(161.5/176); Diego: DI^*A=0.025 6(4.5/176), DI^*B =0.974 4(171.5/176); Kidd: JK^*A=0.485 8(85.5/176), JK^*B=0.514 2(90.5/176); Kell: KP^*A=0.005 7(1/176), KP^*B=0.994 3(175/176); Lutheran: LU^*A=0.005 7(1/176), LU^*B=0.994 3(175/176); Yt: YT^*A=0.002 8(0.5/176), YT^*B=0.997 2(175.5/176). The genotypes of Kell(K+ /k+ ), Scianna and Colton blood groups were KEL^*02 /KEL^*02, SC^*01 /SC^*01 and CO^*A /CO^*B, respectively. The expected frequencies of the combination of type O, RhD negative and other blood group systems were between 1/100 000 to 1/10 000. 【Conclusion】 Among RhD negative blood donors in Hunan, the gene profiles of MNS, Duffy, Domrock, Diego, Kidd, Kell and Lutheran blood group system were polymorphic, and Kell (K+ /k+ ), Colton and Scianna were homozygous. The data of other RBC blood group systems from RhD negative blood donors is of great significance to establish local database of rare blood groups.

Objectives:To investigate the effect of inhibition of long non-coding RNA(lnc RNA)in human metastasis associated lung adenocarcinoma transcript 1(MALAT1)on glycolipitoxicity-induced human umbilical vein endothelial cell dysfunction. Methods:Human umbilical vein endothelial cells were treated with glucose and palmitic acid in vitro to establish the glycolipitoxic endothelial cell models.Following groups were examined:control group,high-glucose and high-fat group,high-glucose and high-fat + non-targeting RAN control group,high-glucose and high-lipid+MALAT1 siRNA group,and high-glucose and high-lipid+MAPK1 siRNA group.RT-qPCR was used to detect the mRNA expression of MALAT1 and MAPK1.Western blot was used to detect the expression levels of autophagy,mitochondrial fusion division,apoptosis,and pathway-related proteins.Immunofluorescence confocal localization was used to detect the fluorescence colocalization of autophagy and lysosome-related proteins.The number of autophagolysosomes in endothelial cells was observed by transmission electron microscopy.Mitochondrial probe staining was used to detect mitochondrial morphology,immunofluorescence was used to detect intracellular reactive oxygen species(ROS)production,flow cytometry was used to detect the apoptosis of cells in each group,cell proliferation and scratch assays were used to detect the proliferation and migration ability of cells in different groups at different time points.The angiogenesis was quantified by counting the number of new blood vessels in each group. Results:Compared with the control group,the expression of lncRNA MALAT1 mRNA and the expression of phosphorylated mito-activated protein kinase 1(p-MAPK1)were upregulated(both P＜0.05)and the expression of phosphorylated mammalian target protein(p-mTOR)was downregulated in the high-glucose and high-fat group and the high-sugar and high-fat control group(all P＜0.01).Compared with the high-glucose and high-fat non-targeting RNA control group,the expressions of microtubule-associated protein 1A/1B-light chain 3(LC3)and p62 were downregulated(P＜0.01,P＜0.05),LC3 and lysosome-associated membrane protein 2(LAMP2)protein co-localized positive fluorescence particles were increased(both P＜0.01),number of lysosomes were decreased,the expression of ROS was decreased(P＜0.01),the expression level of mitochondrial fusion protein optic nerve atrophin 1(OPA1)was increased(P＜0.05),the expressions of cleaved caspase-3 and BCL-2-related X protein(BAX)were decreased and BCL-2 was increased(all P＜0.05),cell proliferation,migration,and tube-forming ability were increased(all P＜0.01),and the expression of p-MAPK1 was decreased(P＜0.05)and p-mTOR expression was increased(both P＜0.05)in the high-glucose and high-lipid+si-MALAT1 group.Compared with the high-glucose and high-fat non-targeting RNA control group,the expression of p-MAPK1 in endothelial cells was decreased and the expression of p-mTOR was increased in the high-glucose and high-lipid+si-MAPK1 group(both P＜0.01). Conclusions:Inhibition of lncRNA MALAT1 expression can reduce the level of mitophagy in glycolipidotoxic environments,reduce apoptosis of endothelial cells and improve endothelial cell function,which may be related to the regulation of MAPK1/mTOR signaling pathway.