Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.

ObjectiveTo investigate the efficacy of acupuncture point injection for fine motor functions in children with cerebral palsy.Methods57 children with cerebral palsy were treated by acupuncture point injection combined with occupational therapy. The efficacy was evaluated by the Peabody Developmental Motor Scale-Fine Motor (PDMS-FM).ResultsThe fine motor quotient (FMQ) of 34 children (59.7%) elevated. The raw scores of grasping subtest (Gr) elevated in 45 children (78.9%) and the raw scores of visual-motor integration (VI) elevated in 53 children (93.0%).ConclusionThe efficacy of acupuncture point injection combined with occupational therapy is significant in children with cerebral palsy.

BACKGROUNDCystic fibrosis (CF) is rare in Chinese. We investigated the mutations in the gene of cystic fibrosis transmembrane conductance regulator (CFTR) in a Chinese CF patient and reviewed the clinical features, gene mutations in Chinese CF cases.

METHODSBlood samples were collected from a previously reported CF girl and her parents. The 24 coding exons of CFTR of the proband were amplified and sequenced.

RESULTSA Chinese girl of 16 years old was diagnosed as CF at the age of 14. She had recurrent productive cough with bronchiectasis in bilateral upper lobes, parasinusitis and otitis media, but without pancreatic involvement. Her sweat chloride was (108.9 +/- 3.3) mmol/L. A heterozygous novel missense mutation of 699 C --> A which results in the amino acid change of N189K was identified in exon 5. In addition, a heterozygous 3821 - 3823 delT mutation in exon 19 was found in CFTR. The mutation 699C --> A was inherited from her father, and the 3821 - 3823 delT mutation was from her mother. Twenty patients with CF in Chinese reported from 1974 to 2004 were also reviewed. DelF508 mutation was not found in the nine cases whose CFTR mutations were analyzed.

CONCLUSIONSThe CF proband carries two heterozygous mutations (699C --> A and 3821 - 3823 delT) in CFTR. 699C --> A mutation is a novel mutation which is not reported previously. Review of reported Chinese cases suggests that the genotype of Chinese CF may be different from those of white cases. More studies are needed to understand the spectra of CFTR and clinical CF features in Chinese.

Adolescent ; Cystic Fibrosis ; complications ; genetics ; Cystic Fibrosis Transmembrane Conductance Regulator ; genetics ; Exocrine Pancreatic Insufficiency ; etiology ; Female ; Humans ; Mutation, Missense ; Respiratory Tract Diseases ; etiology

Objective:To explore the role of parental origin verification in chromosomal microarray analysis (CMA) on the determination of the clinical significance of copy number variations (CNVs).Methods:This retrospective study collected clinical information from 73 core families who underwent prenatal diagnosis at Peking University First Hospital from November 2017 to December 2019. Indications for prenatal diagnosis included ultrasound abnormality in 54 cases (including 12 with thickened nuchal translucency (≥2.5 mm), four with fetal growth restriction, seven with abnormal pregnancy history, and 31 with isolated ultrasound abnormality), NIPT indicated high-risk in four cases, advanced age in nine cases, abnormal pregnancy history alone in three cases, intrauterine death in two cases and one with maternal mental retardation. Genomic DNA of amniotic fluid sample, chorionic villi, cord blood, fetal tissues, and fetal heart blood were extracted using genomic DNA extraction kit. The CNVs of prenatal samples in 73 subjects were analyzed using array-based comparative genomic hybridization (array-CGH) analysis and single nucleotide polymorphism array (SNP-array). Peripheral blood DNA of the couples, and relevant families if necessary, were collected and analyzed in the same way. The results of parental origin detection in CMA were summarized.Results:A total of 76 CNVs were detected in these 73 samples, out of which nine were pathogenic and parental origin detection revealed that six were de novo, two were maternally, and one was paternally inherited; six CNVs were likely pathogenic, including three de novo, two maternally inherited and one paternally inherited; 20 CNVs were variants of uncertain significance, including five paternally inherited, three maternally inherited and 12 de novo; 41 CNVs were likely benign, among which 38 were inherited from parents with normal phenotype. Conclusions:Parental origin verification plays an important role in explaining the clinical significance of detected fetal CNVs and thereby can help to analyze its clinical effect and reproductive risk.

BACKGROUNDTong-xin-luo capsule (TXL), used as a traditional Chinese herb, offeres a therapeutic potential for treatment of cardiovascular diseases. It has been shown to exert a variety of pharmacological effects, including antihypertensive effects, and is able to improve ventricular remodeling. However, the mechanisms of its action are not completely understood. The aim of this study was to evaluate the molecular mechanisms of Tong-xin-luo capsule on left ventricular remodeling in spontaneously hypertensive rats (SHR).

METHODSSixteen eight-week-old SHRs were randomized into an SHR group (n = 8) and a TXL group (n = 8) that were given Tong-xin-luo capsule (1.5 mg x kg(-1) x d(-1)). Eight Wistar Kyoto (WKY) rats fed with 0.9% NaCl served as the control group (WKY group). Systolic blood pressure (BP), body weight and heart rate were monitored once every two weeks. Ventricular remodeling was detected by histopathological examination. Nuclear factor kappa B P65 (NF-kappaB P65) and peroxisome proliferators activated receptor gamma (PPAR-gamma) protein and phosphorylated inhibitor kappa alpha (IkappaBalpha) protein were detected by immunohistochemistry and western blot respectively. The physical interaction of the P65-P50 heterodimer with IkappaBalpha and NF-kappaB were measured by co-immunoprecipitation. PPAR-gamma mRNA, collagen I mRNA and collagen III mRNA were measured by real-time PCR.

RESULTSTXL inhibited NF-kappaB P65 expression and ventricular remodeling and suppressed the activation of NF-kappaB compared with the SHR group (P < 0.01, P < 0.05). TXL reduced IkappaBalpha phosphorylation, increased expression of PPAR-gamma protein and enhanced the physical interaction of the P65-P50 heterodimer with IkappaBalpha. The mRNA expression of PPAR-gamma was enhanced but the mRNA expression of collagen I mRNA and collagen I mRNA were suppressed by TXL.

CONCLUSIONSIn spontaneously hypertensive rats, TXL could inhibit ventricular remodeling induced by hypertension, and the inhibitory effect might be associated with the process of TXL increasing the expression of PPAR-gamma that could result in the inhibition of the activation of NF-kappaB.

Animals ; Collagen Type I ; genetics ; Collagen Type III ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Hypertension ; drug therapy ; pathology ; Male ; NF-kappa B ; antagonists & inhibitors ; PPAR gamma ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Inbred SHR ; Ventricular Function, Left ; drug effects ; Ventricular Remodeling ; drug effects

OBJECTIVETo study the expression and significance of GCS gene in human pancreatic cancer cell line SW1990 and its drug-resistant sublines.

METHODSSW1990 and its drug-resistant sublines, SW1990/FU, SW1990/ADM and SW1990/GEM were cultured in vitro. CCK-8 (Cell Counting kit-8) was used to detect the drug resistance of the sublines. Relative quantitation of GCS mRNA expression was evaluated by real-time PCR and Western blot was adopted to evaluate the expression of GCS protein.

RESULTSThe drug resistance indexes of SW1990/FU, SW1990/ADM and SW1990/GEM were 339.7, 11.9 and 56.6, respectively. The GCS mRNA and protein were expressed in all SW1990 and its drug-resistant sublines. There was a higher expression of GCS mRNA in all the sublines and a significant difference of GCS protein expression was detected in SW1990/ADM and SW1990/GEM compared with SW1990.

CONCLUSIONSGCS gene is expressed in SW1990 and its drug-resistance sublines. The high expression of GCS protein in SW1990/ADM and SW1990/GEM might be one reason of resistance to ADM and GEM in the sublines.

Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Gene Expression ; Glucosyltransferases ; biosynthesis ; genetics ; Humans ; Immunoblotting ; Pancreatic Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Shigella flexneri is an infectious pathogen that causes dysentery to human, which remains a serious threat to public health, particularly in developing countries. In this study, the global protein expression patterns of S. flexneri during transition from exponential growth to stationary phase in vitro were analyzed by using 2-D PAGE combined with MALDI-TOF MS. In a time-course experiment with five time points, the relative abundance of 49 protein spots varied significantly. Interestingly, a putative outer membrane protein YciD (OmpW) was almost not detected in the exponential growth phase but became one of the most abundant proteins in the whole stationary-phase proteome. Some proteins regulated by the global regulator FNR were also significantly induced (such as AnsB, AspA, FrdAB, and KatG) or repressed (such as AceEF, OmpX, SodA, and SucAB) during the growth phase transition. These proteins may be the key effectors of the bacterial cell cycle or play important roles in the cellular maintenance and stress responses. Our expression profile data provide valuable information for the study of bacterial physiology and form the basis for future proteomic analyses of this pathogen.
Bacterial Proteins ; analysis ; Computational Biology ; Electrophoresis, Gel, Two-Dimensional ; Gene Expression Profiling ; methods ; Kinetics ; Peptide Mapping ; Proteome ; analysis ; Proteomics ; methods ; Shigella flexneri ; growth & development ; metabolism ; pathogenicity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Temperature ; Trypsin ; pharmacology ; Virulence

OBJECTIVETo conduct a molecular epidemiological survey on the mitochondrial DNA C1494T mutation in non-syndromic hearing loss patients in Chinese population.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to screen the mitochondrial DNA 12S rRNA C1494T mutation in 20 patients with aminoglycoside antibiotic induced hearing loss, 136 sporadic non-syndromic hearing loss patients and 50 probands of pedigrees with non-syndromic hearing loss.

RESULTSThe C1494T mutation did not appear in all cases except for the positive control.

CONCLUSIONIncidence of mitochondrial DNA C1494T mutation is much lower than that of mitochondrial DNA A1555G mutation in non-syndromic hearing loss of Chinese population. Mitochondrial DNA C1494T mutation may be a rare variation in non-syndromic hearing loss and is not the main cause of aminoglycoside antibiotic induced-deafness.

Adolescent ; Aminoglycosides ; adverse effects ; Anti-Bacterial Agents ; adverse effects ; Asian Continental Ancestry Group ; genetics ; Child ; China ; DNA, Mitochondrial ; genetics ; Female ; Hearing Loss ; chemically induced ; ethnology ; genetics ; Humans ; Male ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA, Ribosomal ; genetics

Adenocarcinoma ; diagnosis ; surgery ; Helicobacter Infections ; diagnosis ; Humans ; Lymphoma, B-Cell, Marginal Zone ; diagnosis ; surgery ; Male ; Middle Aged ; Stomach Neoplasms ; diagnosis ; surgery

This study was conducted to investigate the effect of N, N-diethyldithiocarbamate (DEDTC) on the changes of inflammatory cytokines after focal cerebral ischemia-reperfusion injury in rats and to explore the potential mechanism. Two hundred Sprague Dawley male rats were randomly divided into sham group, middle cerebral artery occlusion (MCAO) group and DEDTC (Zn chelator) treated group. MCAO model was established by the suture method. Rats were sacrificed at 6, 12 and 24 h after reperfusion. 2, 3, 5-Triphenyltetrazolium chloride (TTC) was conducted to measure the brain infarct volume. Newport Green was adopted to detect the chelatable zinc in the cerebral penumbra. Enzyme linked immunosorbent assay (ELISA) was performed to determine the release of TNF-α and IL-6. Furthermore Western blot was used to analyze the expression of the PI3K/Akt/NF-κB signaling pathway. The results showed that DEDTC resulted in a significant reduction of brain infarct volume and an obvious improvement of neurological function compared to the model group. DEDTC also decreased the release of inflammatory cytokines such as TNF-α and IL-6. The activation of PI3K/Akt/NF-κB signaling pathway induced by I/R injury was drastically inhibited by the treatment with DEDTC. In conclusion, DEDTC could protect the brain against ischemic injury induced by MCAO, which might be relevant to the inhibition of PI3K/Akt/NF-κB signaling pathway, and the decreased release of inflammatory cytokines.